CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of United States Provisional Application Serial
Number
61/620,512 filed April 5, 2012, United States Provisional Application Serial Number
61/763,981 filed February 13, 2013, United States Provisional Application Serial Number
61/763,985 filed February 13, 2013, United States Provisional Application Serial Number
61/764,467 filed February 13, 2013, and United States Provisional Application Serial Number
61/764,496 filed February 13, 2013.
BACKGROUND OF THE INVENTION
[0002] Tissue growth, regeneration and repair are often necessary to restore function and
reconstruct the morphology of the tissue, for example, as a result of exposure to
trauma, neoplasia, abnormal tissue growth, aging, and others.
[0003] Synthetic materials have also used as a substrate for promoting ex-vivo tissue assembly
and repair, and similarly for restoring and reconstructing different tissues, for
example for bone, for many years, with mixed success. Another possibility is autologous
tissue grafting, although the supply of autologous tissue is limited and its collection
may be painful, with the risk of infection, hemorrhage, cosmetic disability, nerve
damage, and loss of function. In addition, significant morbidity is associated with
autograft harvest sites. These problems may be overcome by engineering tissue using
solid substrates made of synthetic or natural biomaterials that promote the adhesion,
migration, proliferation, and differentiation of stem cells, for example, mesenchymal
stem cells (MSCs).
[0004] Many diseases and conditions whose treatment is sought would benefit from the ability
to promote cell and tissue growth in a site-specific manner, promoting growth and
incorporation of new tissue within a damaged or diseased site.
[0005] In bone and cartilage applications, the immediate microenvironment and the three-dimensional
(3D) organization are important factors in differentiation in general and particularly
in chondrogenic and osteogenic differentiation.
[0006] Some bone tissue engineering scaffolds consists of natural polymers, such as collagen,
alginate, hyaluronic acid, and chitosan. Natural materials offer the advantages of
specific cell interaction, easy seeding of cells because of their hydrophilic interactions,
low toxicity and low chronic inflammatory response. However, these scaffolds often
are mechanically unstable and do not readily contribute to the creation of tissue
structures with a specific predefined shape for transplantation. To obtain mechanical
strength, chemical modification is required, which may lead to toxicity.
[0007] Defects and degeneration of the articular cartilage surfaces of joints causes pain
and stiffness. Damage to cartilage which protects joints can result from either physical
injury as a result of trauma, sports or repetitive stresses (e.g., osteochondral fracture,
secondary damage due to cruciate ligament injury) or from disease (e.g. osteoarthritis,
rheumatoid arthritis, aseptic necrosis, osteochondritis dissecans).
[0008] Osteoarthritis (OA) results from general wear and tear of joints, most notably hip
and knee joints. Osteoarthritis is common in the elderly but, in fact, by age 40 most
individuals have some osteoarthitic changes in their weight bearing joints. Another
emerging trend increasing the prevalence of osteoarthritis is the rise in obesity.
The CDC estimates that 30% of American adults (or 60 million people) are obese. Obese
adults are 4 times more likely to develop knee OA than normal weight adults Rheumatoid
arthritis is an inflammatory condition which results in the destruction of cartilage.
It is thought to be, at least in part, an autoimmune disease with sufferers having
a genetic predisposition to the disease.
[0009] Orthopedic prevention and repair of damaged joints is a significant burden on the
medical profession both in terms of expense and time spent treating patients. In part,
this is because cartilage does not possess the capacity for self-repair. Attempts
to re-grow hyaline cartilage for repair of cartilage defects remain unsuccessful.
Orthopedic surgery is available in order to repair defects and prevent articular damage
in an effort to forestall serious degenerative changes in a joint. The use of surgical
techniques often requires the removal and donation of healthy tissue to replace the
damaged or diseased tissue. Techniques utilizing donated tissue from autografts, allografts,
or xenografts are wholly unsatisfactory as autografts add additional trauma to a subject
and allografts and xenografts are limited by immunological reactivity to the host
subject and possible transfer of infective agents. Surgical attempts to utilize materials
other than human or animal tissue for cartilage regeneration have been unsuccessful.
[0010] WO 2010/146575 A1 discloses aragonite- and calcite-based scaffolds for the repair, regeneration, enhancement
of formation or a combination thereof of cartilage and/or bone, which scaffolds comprise
at least two phases, wherein each phase differs in terms of its chemical content,
or structure, kits comprising the same, processes for producing solid aragonite or
calcite scaffolds and methods of use thereof.
[0011] An ideal material which restores tissue function and facilitates reconstruction of
the morphology of such tissue is as yet, lacking.
SUMMARY OF THE INVENTION
[0012] The scope of this invention is defined by the claims. Embodiments in the description
relating to methods of treatment are not covered by the claims. Any "embodiment" or
"example" which is disclosed in the description but is not covered by the claims should
be considered as presented for illustrative purpose only.
[0013] This invention provides a solid substrate for tissue repair, said solid substrate
consisting essentially of three phases wherein:
- a first phase of said three phases comprised substantially of a first biocompatible
polymer, which first phase is characterized by being comprised of an elastic material
which is substantially less rigid in structure than that of said second and third
phases of said three phases;
- a second phase of said three phases comprises a marine organism skeletal derivative
-based solid substrate further comprising a second biocompatible polymer, wherein
said second phase further comprises a series of hollows along a longitudinal axis
in said second phase, and wherein said second biocompatible polymer is substantially
located within said series of hollows; and
- a third phase of said three phases comprises a marine organism skeletal derivative
-based solid substrate, optionally comprising a series of hollows along a longitudinal
axis in said third phase,
wherein said marine organism skeletal derivative-based solid substrate is a coral
or coral derivative, wherein said coral or coral derivative is aragonite, calcite,
hydroxyapatite, mixtures thereof, or other polymorphs of the same and wherein said
marine organism skeletal derivative is characterized by a specific fluid uptake capacity
value of at least 75%, which specific fluid uptake capacity value is determined by
establishing a spontaneous fluid uptake value divided by a total fluid uptake value,
wherein said fluid is a protein-containing, salt-containing or carbohydrate containing
solution, or wherein said fluid is a hydrophilic biologic fluid, wherein said biologic
fluid is autologous with respect to a cell or tissue of a subject when said solid
substrate is contacted with a cell or tissue of said subject or wherein said fluid
is water, or wherein said marine organism skeletal derivative is characterized by
having a contact angle value of less than 60 degrees, when in contact with a fluid,
wherein said fluid is a protein-containing, salt-containing or carbohydrate containing
solution, or wherein said fluid is a hydrophilic biologic fluid, wherein said biologic
fluid comprises cells or tissue, or wherein said fluid is water.
[0014] In some embodiments, according to this aspect, the first biocompatible polymer is
in the form of a hydrogel.
[0015] In some embodiments, said first phase is elastic following wetting in situ.
[0016] In some embodiments, according to this aspect, the first or second biocompatible
polymer or a combination thereof comprises collagen, cross-linked collagen, chitosan,
elastin, silk, aliginate, fibrin, platelet rich plasma, a glycosaminoglycan, or combinations
thereof. In some embodiments, according to this aspect, the glycosaminoglycan is hyaluronic
acid, sodium hyaluronate, or a cross linked hyaluronic acid or a combination thereof.
In some embodiments, according to this aspect, the alginate may comprise calcium alginate,
cross linked calcium alginate or a combination thereof. In some embodiments, according
to this aspect, the chitosan may comprise cross linked chitosan.
[0017] In some embodiments, a third, fourth, or any number of additional biocompatible polymers
may be incorporated within the solid substrates of this invention.
[0018] In some embodiments, according to this aspect, the solid substrate further comprises
at least one cell population suspended therein or seeded thereon. In some embodiments,
according to this aspect, the at least one cell population is a population of chondrocytes
or progenitor cells therefor.
[0019] In some embodiments, the solid substrate is isolated from a Porites species, a Goniopora,
a Millepora species or an Acropora species.
[0020] In some embodiments, according to this aspect, the first or second biocompatible
polymer further comprises a cytokine, a growth factor, a chelator, a cell population,
a therapeutic compound, a drug, or any combination thereof. In some embodiments, according
to this aspect, the therapeutic compound or drug comprises an anti-inflammatory compound,
an anti-infective compound, a pro-angiogenic factor or a combination thereof.
[0021] In some embodiments, according to this aspect, the said first and said second phase
are designated for insertion into a region which is proximal to cartilage and said
third phase is designated for insertion into a region which is proximal to subchondral
bone.
[0022] In one alternative, the marine organism skeletal derivative is characterized by a
specific fluid uptake capacity value of at least 75%, which specific fluid uptake
capacity value is determined by establishing a spontaneous fluid uptake value divided
by a total fluid uptake value.
[0023] In another alternative, the marine organism skeletal derivative is characterized
by having a contact angle value of less than 60 degrees, when in contact with a fluid.
[0024] In one alternative, according to these aspects, the fluid is a protein-containing,
salt-containing or carbohydrate containing solution. In another alternative, the fluid
is a biologic fluid. In some embodiments, the biologic fluid is autologous with respect
to a cell or tissue of a subject when said solid substrate is contacted with a cell
or tissue of said subject. In another alternative, the fluid is water.
[0025] Also disclosed is a method of inducing or enhancing repair, regeneration or enhancement
of formation of cartilage, bone or a combination thereof, said method comprising implanting
in a subject, a solid substrate as herein described (and in accordance with any permutation
thus described) within a site in need of repair, regeneration or enhancement of formation
of cartilage, bone or a combination thereof.
[0026] In some disclosures, the method comprises rendering the first phase more elastic
following wetting of the first phase as a consequence of or during the implantation
of the solid substrate.
[0027] The invention provides also a kit for repair of cartilage comprising:
- at least one first biocompatible polymer which is an elastic material which is less
rigid in structure than that of a first and a second phase, and
- a solid substrate which substrate comprises the first phase comprising a marine organism
skeletal derivative -based solid substrate comprising a series of hollows along a
longitudinal axis in said first phase, and a second biocompatible polymer located
within said series of hollows; and
the second phase comprising a marine organism skeletal derivative -based solid substrate,
wherein said marine organism skeletal derivative-based solid substrate is a coral
or coral derivative, wherein said coral or coral derivative is aragonite, calcite,
hydroxyapatite, mixtures thereof, or other polymorphs of the same;
and wherein the kit is provided with instructions and appropriate tools for applying
the second biocompatible polymer to the solid substrate such that some of the second
biocompatible polymer is substantially located within the series of hollows in the
substrate and such that some of the first biocompatible polymer creates a separate
phase at a terminus of the solid substrate and the first biocompatible polymer is
added to said solid substrate in situ, and wherein said marine organism skeletal derivative
is characterized by a specific fluid uptake capacity value of at least 75%, which
specific fluid uptake capacity value is determined by establishing a spontaneous fluid
uptake value divided by a total fluid uptake value, wherein said fluid is a protein-containing,
salt-containing or carbohydrate containing solution, or wherein said fluid is a hydrophilic
biologic fluid, wherein said biologic fluid is autologous with respect to a cell or
tissue of a subject when said solid substrate is contacted with a cell or tissue of
said subject or wherein said fluid is water, or wherein said marine organism skeletal
derivative is characterized by having a contact angle value of less than 60 degrees,
when in contact with a fluid, wherein said fluid is a protein-containing, salt-containing
or carbohydrate containing solution, or wherein said fluid is a hydrophilic biologic
fluid, wherein said biologic fluid comprises cells or tissue, or wherein said fluid
is water.
[0028] In some embodiments, the solid substrate comprises a third phase created when said
second biocompatible polymer is applied, which second biocompatible polymer is of
the same material as said first biocompatible polymer, and wherein said first phase
is assembled, by applying said first biocompatible polymer at the same time as applying
said second biocompatible polymer.
[0029] In some disclosures, the first and third phases are assembled during or following
the implantation of the solid substrate.
[0030] In some disclosures, the method comprises exposing a site of cartilage repair, and
optionally exposing bone tissue located proximally to the site of cartilage repair
in the subject prior to implanting the solid substrate.
[0031] In some embodiments, this invention provides for the use of a solid substrate as
herein described (in any embodied permutation as described herein) for inducing or
enhancing repair, regeneration or enhancement of formation of cartilage, bone or a
combination thereof.
[0032] Where number ranges are given in this document, endpoints are included within the
range. Furthermore, it is to be understood that unless otherwise indicated or otherwise
evident from the context and understanding of one of ordinary skill in the art, values
that are expressed as ranges can assume any specific value or sub-range within the
stated ranges, optionally including or excluding either or both endpoints, in different
embodiments of the invention, to the tenth of the unit of the lower limit of the range,
unless the context clearly dictates otherwise. Where a percentage is recited in reference
to a value that intrinsically has units that are whole numbers, any resulting fraction
may be rounded to the nearest whole number.
BRIEF DESCRIPTION OF THE DRAWINGS
[0033]
Figure 1 schematically depicts an embodiment of a solid substrate/implants of this
invention. According to this aspect, a first phase 1-10 is located most apically and comprises a first biocompatible polymer, and a second
phase 1-20 comprises aragonite throughout which are a series of holes or voids along a longitudinal
axis which are impregnated or accumulated therewithin is a second biocompatible polymer,
such as hyaluronic acid and a third phase 1-30 comprising only aragonite.
Figure 2 schematically depicts the embodied solid substrate of Figure 1, cut along
a longitudinal axis, in order to better view the series of voids 2-40 along the longitudinal axis.
Figure 3 schematically depicts the embodied scaffold of Figure 1, with the porosity
of the aragonite-containing elements emphasized, as well as depicting the presence
of a second biocompatible polymer 3-45 within the voids in the second phase 3-20. The first 3-10 and third phases 3-30, respectively, are depicted, as well.
Figure 4 provides photographs of an embodied scaffold of this invention, showing top/front
and side views. The first 4-10 and third phases 4-30, respectively, are evident, with the second phase shown 4-20, more clearly depicted in the depiction in Figure 3.
Figure 5 schematically depicts the insertion of an embodied solid substrate within
a defect site spanning bone 5-70 and cartilage 5-60. The first phase 5-10 and second phase 5-20 are both located within the cartilage layer, whereas the third phase 5-30 is located within the bone component. The voids within the second layer 5-40 are depicted, as well, but in this aspect, penetration of the biocompatible polymer
therewithin is not readily seen. The diagram depicts the incorporation of appropriate
cells within the implant 5-50 responsible for tissue regeneration and repair at the implantation site, as well.
DETAILED DESCRIPTION OF THE PRESENT INVENTION
[0034] This invention provides,
inter alia, solid substrates, for repair of cartilage tissue in a subject. This invention further
provides a kit for repair of cartilage tissue in a subject.
[0035] This invention provides,
inter alia, optimized solid substrates for promoting cell or tissue growth or restored function
and processes for producing the same.
[0036] Coral, which is comprised of CaCO
3 in the crystalline form of aragonite or calcite has the advantage of supporting fast
cellular invasion, adherence, proliferation and differentiation of mesenchymal stem
cells into cartilage and/or bone tissue.
[0037] Three-dimensional (3-D) solid substrates attract mesenchymal stem cells from surrounding
or proximally located tissue and promote blood vessel formation to a site of cartilage
repair. Such solid substrates can be used for regeneration, repair and enhancement
of formation of cartilage and/or bone in a subject for the treatment of full-thickness
cartilage defects, partial thickness cartilage defects and/or osteochondral defects.
[0038] The solid substrates of this invention will comprise a marine organism skeletal derivative
-based material.
[0039] The term "marine organism skeletal derivative -based material" refers to a solid
piece or ground material derived from a marine organism, and from a skeletal component
of the organism, such as an exoskeleton of the same.
[0040] More specifically, the term "marine organism skeletal derivative -based material"
refers to a coralline-based material. Coral, which is comprised of CaCO
3 in the crystalline form of aragonite or calcite has been shown to possess the advantage
of supporting fast cellular invasion, adherence and proliferation. Coral has been
shown to be an effective substrate for facilitation of the adherence, proliferation
and differentiation of mesenchymal stem cells, and ultimate incorporation into cartilage
and/or bone tissue. Coral has also been shown to serve as an excellent substrate for
promoting adherence and proliferation of a number of other cell types, serving as
an excellent support for cell and tissue growth.
[0041] The terms "coral" and "aragonite" and "calcite" may be used interchangeably herein.
[0042] More specifically, the term "marine organism skeletal derivative -based material"
refers to a coral or coral derivative.
[0043] In some embodiments, the solid substrate contains ground particles derived from coral,
suspended in a biocompatible matrix. In some embodiments, the biocompatible matrix
is a hydrogel.
[0044] In some embodiments, reference to an "implant" or "plug" or "solid substrate", as
used herein refers to any embodiment or combined embodiments as herein described with
regard to the solid substrates and to be considered as being included in the described
aspect of this invention. For example, reference to a "solid substrate" as used herein,
is to be understood to refer to any embodiment of a solid substrate as described herein
being applicable for the indicated purpose or containing the indicated attribute,
etc.
[0045] In one embodiment, "solid substrate" refers to a shaped platform used for cell and/or
tissue repair and/or restored function, wherein the shaped platform provides a site
for such repair and/or restored function. In one embodiment, the solid substrate is
a temporary platform. In one embodiment, "temporary platform" refers to a natural
degradation of a coral of this invention that occurs over time during such repair,
wherein the natural fully or partially degradation of the coral may results in a change
of solid substrate shape over time and/or change in solid substrate size over time.
[0046] It will be appreciated that different species of coral vary in terms of their average
pore diameter and pore volume and the invention contemplates use of any such coral
as a starting material for the preparation of the solid substrates as herein described.
As used herein, the term "pore volume" refers to volume or open spaces inside the
porous scaffolding of this invention. Pore volume is determined by any means known
in the art. Porosity can be calculated by standard methods, an example of which is
provided further hereinbelow, see for example,
Karageorgiou V, Kaplan D. (2005) "Porosity of 3D biomaterial scaffolds and osteogenesis"
Biomaterials.;26(27):5474-91.
[0047] It will be appreciated that the term "coral" will refer to a starting material from
which aragonite, calcium carbonate, calcite, or hydroxyapatite etc. may be isolated.
[0048] The coralline-based or calcite-based, etc., solid substrates of this invention may
also be used for regeneration, repair and enhancement of formation of bone in a subject,
for the treatment of a bone condition, disease or disorder.
[0049] This invention provides the unexpected application of the described solid substrates
for use in a three-phase arrangement, either inserted as a tri-phasic scaffolding
material, or inserted as a bi-phasic solid substrate, whereby the third phase is assembled
in situ. Such solid substrates are uniquely and superiorly useful in cartilage and/or
bone regeneration, repair and enhancement of formation and moreover, that such solid
substrates can be prepared and inserted specifically and optimally within cartilage
and/or bone in a subject in need thereof, for methods of cartilage and/or bone regeneration,
repair and enhancement of formation.
[0050] In particular, this invention provides the unexpected application that cartilage
and/or bone regeneration, repair and enhancement of formation is optimal when the
coral scaffolding consists essentially of three phases wherein:
- a first phase of said three phases comprised substantially of a first biocompatible
polymer, which first phase is characterized by being comprised of an elastic material,
and which material is substantially less rigid in structure than that of said second
and third phases of said three phases;
- a second phase of said three phases comprises s marine organism skeletal derivative
-based solid substrate comprising a second biocompatible polymer and said second phase
further comprises a series of hollows along a longitudinal axis in said second phase,
wherein said biocompatible polymer is substantially located within said series of
hollows; and
- a third phase of said three phases comprises a marine organism skeletal derivative
-based solid substrate, optionally comprising a series of hollows along a longitudinal
axis in said third phase.
[0051] In particular, this invention provides the unexpected advantage in terms of greater
incorporation of the solid substrate within the newly developed cartilage, and reduced
inflammation, when the solid substrates are structured such as those herein described.
[0052] Another advantage to the solid substrates according to this aspect, is the presence
of pre-drilled channels or longitudinally placed holes within the phase containing
the biocompatible polymer such as hyaluronic acid, which holes are impregnated with
the biocompatible polymer such as hylauronic acid and serve as a reservoirs for the
biocompatible polymer such as hyaluronic acid in a phase located within a region in
need of cartilage repair. Localization of the biocompatible polymer such as hyaluronic
acid allows for greater direction of migrating progenitor cells throughout the phase
of this solid substrate to stimulate cartilage regeneration and repair. In some embodiments,
the channels comprising concentrated biocompatible polymer such as hyaluronic acid
within the voids along the longitudinal axis of the phase of the solid substrate provide
a chemotactic guide for recruited cells involved in chondrogenesis, and/or in some
embodiments, influence local recruitment and differentiation of the chondrogenic population
of cells migrating thereto. In some embodiments, the channels comprising concentrated
biocompatible polymer such as hyaluronic acid within the voids along the longitudinal
axis of the phase of the solid substrate contribute to cartilage matrix homeostasis.
[0053] Unexpectedly, it was found that apical placement of a phase consisting essentially
of the first biocompatible polymer, and positioning of the same within the site of
repair, or positioning the same slightly under, flush or slightly over the upper limit
of the site of defect provides for greater incorporation within the defect site and
reduced inflammation associated therewith.
[0054] According to this aspect, the porosity and greater rigidity of the third phase as
compared to the first is more suited for insertion within bone and provides a support,
for the repair of osteochondral defects. The solid substrates of this invention are
therefore, in some embodiments, ideally suited for incorporation within a defect site
that spans two different types of tissue, i.e. bone and cartilage.
[0055] In some embodiments, according to this aspect, the first phase has a height of between
0.1-4 mm, or in some embodiments, 0.1-3 mm, or in some embodiments, 0.1-1 mm, or in
some embodiments, 0.5-3 mm, or in some embodiments, 0.5-4 mm, or in some embodiments,
0.5-2 mm, or in some embodiments, 1-2 mm, or in some embodiments, 1-3 mm, or in some
embodiments, 1-4 mm, or in some embodiments, 0.01-0.5mm, or in some embodiments, 0.03-0.1mm,
or in some embodiments, 0.05-0.15mm.
[0056] In some embodiments, the first and second biocompatible polymer, are hydrophilic,
and when synovial fluid comes into contact therewith at the apical layer above the
solid substrate, or when saline comes into contact therewith during the implantation
procedure, the implant absorbs the fluid and reverts to a hydrogel, as opposed to
the pre-implantation dehydrated/dessicated state. This reversion provides mechanical
protection at the site of implantation, in some embodiments.
[0057] In some embodiments, the exterior layer, when "reconstituted" as described following
implantation, may elute from the solid substrate into the surrounding site and thereby
participate in the stimulation or enhancement of repair at the site, including
inter alia, serving as a chemoattractant for cells involved in the repair process.
[0058] In some disclosures, a bi-phasic implant consisting of the second and third phases
as herein described is inserted within a defect site and the first phase is applied
apically above the implanted scaffolding, creating or constituting in situ assembly
of the three-phase implant.
[0059] It will be appreciated by the artisan, that reference to the terms "first", "second"
and "third", in particular, as relating to the described phases within the implants
of this invention, are to designate the presence of the one, two or three elements,
and are to be defined by their constituents.
[0060] Thus, in one embodiment, this invention provides a method of inducing or enhancing
repair, regeneration or enhancement of formation of cartilage, bone or a combination
thereof, said method comprising implanting in a subject, a solid substrate this invention
(including any embodiment with respect thereto as herein described), within a site
in need of repair, regeneration or enhancement of formation of cartilage, bone or
a combination thereof. In some embodiments, the method comprises rendering said first
phase more elastic following wetting of said first phase as a consequence of or during
said implanting.
[0061] Thus, this disclosure provides a method of inducing or enhancing repair, regeneration
or enhancement of formation of cartilage, bone or a combination thereof, said method
comprising implanting within a site in need of repair, regeneration or enhancement
of formation of cartilage, bone or a combination thereof in a subject, a solid substrate
comprising:
- a first phase comprising a marine organism skeletal derivative -based solid substrate
comprising a first biocompatible polymer and said first phase further comprises a
series of hollows along a longitudinal axis in said first phase, wherein said first
biocompatible polymer is substantially located within said series of hollows; and;
and
- a second phase comprising a marine organism skeletal derivative -based solid substrate,
optionally comprising a series of hollows along a longitudinal axis in said second
phase;
whereby said implanting places said solid substrate within said site in need of repair,
and
- applying a second biocompatible polymer to a region above said first phase to be slightly
less than, be flush with or be slightly above an upper limit of said site in need
of repair.
[0062] In some embodiments, the solid substrate comprises a third phase created when said
second biocompatible polymer is applied, which second biocompatible polymer is of
the same material as said first biocompatible polymer, and wherein said first phase
is assembled, by applying said first biocompatible polymer at the same time as applying
said second biocompatible polymer. In some embodiments, the first and third phases
are assembled during or following implantation.
[0063] The phase comprising a biocompatible polymer, and further comprising a series of
hollows along a longitudinal axis in said phase, wherein the biocompatible polymer
is substantially located within the series of hollows, is typically referred to herein
as the second phase. Such second phase will comprise a series of longitudinal holes,
which longitudinal holes may range from 15-60 holes placed throughout the phase along
a longitudinal axis of the implant according to this aspect. In some embodiments,
the holes or enlarged voids will have a diameter ranging from about 250-450 µm. In
some embodiments, the holes or enlarged voids will have a diameter ranging from about
125-650 µm, or in some embodiments, ranging from about 175-550 µm.
[0064] According to this aspect, and in some embodiments, the series of holes or voids may
be incorporated by physical manipulation of the implant, for example, and in some
embodiments, solid aragonite or calcite may be isolated, cleaned and otherwise prepared
as described herein, and a drill may be used to create the series of holes/voids as
herein described. In some embodiments, other means, such as selective dissolution
of the scaffolding material may be accomplished, where the selective dissolution along
a longitudinal axis is accomplished by methods known in the art, including those described
and exemplified herein.
[0065] According to this aspect, the solid substrates further comprise a phase containing
a marine organism skeletal derivative -based solid substrate, such as, for example,
solid coral or biolattice, which has not been further modified to alter the porosity
of the phase, or in some embodiments, may be altered as described further hereinunder
to specifically alter the pore volume or average pore diameter in the phase, whereby
such modifications are substantially uniform throughout the phase. Such phase is typically
referred to herein as the third phase.
[0066] Incorporation of a biocompatible polymer such as hyaluronic acid in the first and
second phase of the implant or in both phases, may be accomplished via any means,
including pressure-driven application, for example, via application under vacuum,
centrifugal force or mechanical pressure. In some embodiments, gravitational force
is sufficient to allow appropriate and relatively homogenous penetration of the hyaluronic
acid to a desired depth of the implant, creating the first phase as herein described.
According to this aspect, in one embodiment, visual inspection of the implant, for
example using the staining with Fast Green/ Safranin O, demonstrates uniform distribution
of the hyaluronic acid through the phase and to a desired depth as a function of the
time and conditions of application.
[0067] According to this aspect, when applying the solid substrates to a site of bone and/or
cartilage repair, or in some embodiments, to a defect site where both bone and cartilage
are affected and in need of repair and/or regeneration, the skilled artisan will appreciate
that the third phase of the solid substrate is inserted within the bone defect site
whereas the first and second phases are inserted within the cartilage defect site
and are ensured to be flush with or slightly below the upper limit of the defect site.
[0068] In some disclosures, such solid substrates may be administered to a subject with
a bone defect in need of repair, wherein access to the bone defect results in the
creation of a defect in the overlying cartilage, and the solid substrates of this
invention allow for the healing of both affected tissues. In other disclosures, such
solid substrates may be administered to a subject with a cartilage defect in need
of repair, wherein optimal insertion of the solid substrate for stimulation of cartilage
repair necessitates anchoring of the solid substrate in the underlying bone, for example,
by creating a minimal void in the underlying bone for insertion of the solid substrate,
and once inserted, the solid substrate facilitates repair of both the overlying cartilage
and underlying bone.
[0069] In other disclosures, such solid substrates may be administered to a subject with
an osteochondral defect, where both bone and cartilage tissue are in need of repair
as part of the pathogenesis of the disorder. The solid substrates according to this
aspect are, in some embodiments, particularly suited for such applications.
[0070] This invention also provides for the unexpected effect that cartilage and/or bone
regeneration, repair and enhancement of formation is optimal when the coral scaffolding
comprises at least two phases, which phases comprise voids, and vary in terms of the
average diameter of the voids within each phase, and/or that cartilage and/or bone
regeneration, repair and enhancement of formation is optimal when the coral scaffolding
comprises at least two phases, which phases vary in terms of their respective pore
volumes (porosity).
[0071] It will be appreciated that the term "coral" will refer to a starting material from
which aragonite and/or calcite may be isolated.
[0072] The present disclosure also provides a solid substrate for inducing or enhancing
cartilage or bone regeneration, repair, enhancement of formation, or a combination
thereof, which solid substrate consists of a solid form of aragonite or calcite and
further comprises :
- at least a first phase, comprising voids having an average diameter ranging from about
60-160 µm; and
- at least a second phase, comprising voids having an average diameter ranging from
about 170-850 µm.
[0073] It will be appreciated that according to this aspect, the term "first phase" and
"second phase" do not apply to a particular order with respect to insertion of the
phase within an osteochondral defect, and either the first phase or the second phase
may be oriented to be proximal to cartilage within a repair site, as opposed to the
prior embodiment of a solid substrate as described hereinabove, wherein the first
phase is inserted proximal to a site of cartilage repair. According to this aspect,
the solid substrate may be further modified to comprise both the indicated void average
diameter, and either phase may further comprise a biocompatible polymer such as hyaluronic
acid and a series of voids or holes along a longitudinal axis of said phase, wherein
the biocompatible polymer such as hyaluronic acid is located substantially within
such series of voids or holes.
[0074] In some embodiments, the term "solid form" with respect to aragonite, refers to solid
aragonite harvested from coral, which aragonite is treated to remove debris, proteins
and other particulate matter, however, such coral-derived materials are not hydrothermically
transformed, nor ground, and resuspended.
[0075] In some embodiments, the coral for use in the preparation of the solid substrates
of this invention may be processed by any means known in the art, for example, as
described in PCT International Application Publication Number
WO 2009/066283, PCT International Application Publication Number
WO 2010/058400, PCT International Application Publication Number
WO 2010/146574 and PCT International Application Publication Number
WO 2010/146574.
[0076] In some embodiments, the coral may be processed according to a process as herein
described.
[0077] In some embodiments, this invention provides a process for the purification of a
coralline-based scaffolding, said process comprising the steps of:
- contacting solid aragonite of a desired size and shape with a solution comprising
an oxidizing agent; and
- washing and drying said solid aragonite
whereby one or each of said steps is conducted under applied negative pressure.
[0078] According to this aspect, and in some embodiments, the applied negative pressure
ranges between about 2.10
4 Pa to 1 Pa (0.2 to 0.00001 Bar) or in some embodiments, the applied negative pressure
ranges between 4.10
4 Pa to 0.01 Pa (0.4 to 0.0000001 Bar)
[0079] According to this aspect, and in some embodiments, the oxidizing agent for use in
the processes of this invention may be any suitable oxidizing agent, which facilitates
the removal of organic debris from coralline-based solid substrates.
[0080] In some embodiments, the oxidizing agent may include, inter alia, potassium nitrate
(KNO3), hypochlorite and other hypohalite compounds, iodine and other halogens, chlorite,
chlorate, perchlorate, permanganate salts, ammonium cerium(IV) nitrate, hexavalent
chromium compounds, pyridinium chlorochromate (PCC), and chromate/dichromate compounds,
peroxide compounds, sulfoxides, persulfuric acid, or nitric acid , acetone, ammonium
peroxydisulfate, 1,4-benzoquinone, N-tert-butylbenzensulfinilmidoyl, chloride, tert-butyl
hydroperoxide, tert-butyl hypochlorite, 3-chloroperoxybenzoic acid, meta-chloroperbenzoic
acid, cumene hydroperoxide, dimethyl sulfoxide, hydrogen peroxide, manganese oxide,
meta-chloroperbenzoic acid, N-methylmorpholine-N-oxide, methyltrioxorhenium (MTO),
oxalyl chloride, N-tert-butylbenzenesulfinimidoyl chloride, oxone, oxygen, ozone,
peracetic acid, periodic acid, peroxy acid, pivaldehyde, potassium permanganate, potassium
peroxydisulfate, potassium peroximonosulfate, 2-propanone, sodium chlorite, sodium
percarbonate, sodium periodate, styrene, trichloroisocyanuric acid (TCCA), 2,2,6,6-tetramethylpiperidinyloxy
TEMPO, tert-butyl hydroperoxide, tert-butyl hypochlorite, tetrabutylammonium peroxydisulphate,
trimethylacetaldehyde. In some embodiments, the oxidizing agent is sodium hypochlorite.
[0081] According to this aspect, and in some embodiments, the process comprises conducting
said contacting under mildly acidic conditions.
[0082] According to this aspect, and in some embodiments, the process comprises subjecting
the solid aragonite to a temperature of at least 275 °C under applied negative pressure.
[0083] According to this aspect of the invention, the process comprises contacting the aragonite
with an oxidizing agent under applied negative pressure, washing and drying the aragonite
applied negative pressure, or both steps are conducted under applied negative pressure.
The applied negative pressure ranges between 2.10
4 Pa to 1 Pa (0.2 to 0.00001 Bar), or in some embodiments, between about 4.10
4 Pa to 0.01 Pa (0.4 to 0.0000001 Bar), according to this aspect of the invention.
[0084] The solid substrates, kits, processes and methods of this invention make use of solid
coralline forms.
[0085] The solid forms or solid substrates of this invention may be of aragonite or calcite
origin.
[0086] In some embodiments, the term "solid form" with respect to calcite refers to calcite
isolated from coral, which calcite is treated to remove debris, proteins and other
particulate matter, however, such materials are not hydrothermically transformed,
nor ground, and resuspended. In some embodiments, the "solid form" calcite refers
to calcite obtained by the preparation of an aragonite solid form, which form is then
converted to calcite by known methods in the art, for example by exposing the form
to high temperature under vacuum.
[0087] Any method for conversion of aragonite to calcite as known in the art may be used
to prepare calcite solid substrates of this invention.
[0088] In some embodiments, coral-based solid substrates of this invention may be converted
to partially or fully into hydroxyapatite by known methods.
[0089] In some embodiments, coral-based solid substrates of this invention may be converted
to partially or fully into hydroxyapatite by known methods.
[0090] In some embodiments, the solid substrates of this invention comprise a series of
voids, and the at least two phases present in the scaffolding of this invention vary
in terms of the average diameter of the voids present in each phase. In some embodiments,
the solid substrate will comprise at least a first phase, comprising voids having
an average diameter ranging from about 60-160 µm. In some embodiments, the first phase
comprises voids having an average diameter ranging from about 60-90 µm, or in some
embodiments, from about 80-130µm, or in some embodiments, from about 120-160 µm.
[0091] In some embodiments, the solid substrate will comprise at least a second phase, comprising
voids having an average diameter ranging from about 170-850 µm. In some embodiments,
the second phase comprises voids having an average diameter ranging from about 170-400
µm, or in some embodiments, from about 250-500 µm, or in some embodiments, from about
450-700 µm or in some embodiments, from about 550-850 µm
[0092] In some embodiments, according to this aspect, the solid substrate further comprises
a third phase, comprising voids having an average diameter ranging from about 150-300
µm and said second phase comprises voids having an average diameter ranging from about
350-850 µm and said third phase is positioned between said first and second phase.
In some embodiments, such at least third phases may be referred to herein interchangeably
as an "intermediate phase".
[0093] In some embodiments, the solid substrate is cylindrical in shape and has a diameter
of about 5-15 mm, and a height of about 5-25 mm. In some embodiments, the solid substrate
has a diameter of about 1-35 mm, and a height of about 1-45 mm, or about 5-40 mm,
and a height of about 5-60 mm, or about 5-15 mm, and a height of about 5-45 mm.
[0094] The average diameter of the voids within the phases of the scaffolding of this invention
may be determined by any means, including digital images analysis, as exemplified
further hereinbelow. In one embodiment, a coral for use in a solid substrate of this
invention comprises an average void diameter appropriate for cell seeding and/or development
of vasculature.
[0095] The solid forms of this invention comprise at least three phases, two of which phases
contain pores, owing to the porous nature of the materials of which the scaffolding
is comprised. In some embodiments, the phases vary in terms of the pore volume (porosity)
in each phase.
[0096] It will be appreciated that different species of coral vary in terms of their average
pore diameter and pore volume and the invention contemplates use of any such coral
as a starting material for the preparation of the solid substrates as herein described.
[0097] By optimizing the specific positioning of a solid substrate the porous crystalline
structure of a coral solid substrates of this invention, described below, is accessible
to beneficial components located within a tissue milieu. For example, the porous crystalline
structure of coral allows in-growth of blood vessels to create a blood supply for
the cartilage that will infiltrate the solid substrate during cartilage repair. By
penetrating into a bone marrow void, mesenchymal stem cells located within the bone
marrow now have access to the exposed surface of the solid substrate. In one embodiment,
the region of the solid substrate penetrating into a bone marrow void attracts mesenchymal
stem cells from the bone marrow and promotes blood vessel formation to the site of
cartilage repair. In one embodiment, the region of the solid substrate penetrating
into a bone marrow void promotes adhesion, proliferation, or differentiation or a
combination thereof, of the mesenchymal stem cells attracted to the solid substrate.
[0098] Thus, it will be apparent to one skilled in the art that the specific positioning
of the solid substrate within a site of cartilage repair arranges the solid substrate
of this invention such that the solid substrate is most effective for cartilage and
bone repair.
[0099] In some embodiments, the improved solid substrates of this invention allow for greater
solid substrate incorporation within newly developed cartilage, reduced inflammation
at the implant site or a combination thereof.
[0100] In some embodiments, the solid substrate approximates the form of a cylinder, cone,
tac, pin, screw, rectangular bar, plate, disc, pyramid, granule, ball or cube.
[0101] In some embodiments, the solid substrates of this invention may be used in conjunction
with other known and/or available materials for stimulating/enhancing bone and/or
cartilage repair. In some embodiments, the solid substrates of this invention may
be utilized to affix additional solid substrates, for example for use in whole joint
repair or ligament repair, or other connector tissue repair.
[0102] In some embodiments, the solid substrates of this invention may be used for example,
as a pin, in conjunction with other solid substrates for bone repair or regeneration,
etc. It is to be understood that any use of the solid substrates of this invention,
alone or in conjunction with other appropriate materials, for the treatment, repair
or stimulation of growth of bone and/or cartilage is to be considered as part of this
invention
[0103] It will be appreciated that the solid substrates of this invention may be of any
suitable shape or size to accommodate its application in accordance with the methods
of this invention. For example, and in some embodiments, for applications of the solid
substrates of this invention within long bones of a subject, the dimensions of the
solid substrate will be scaled to approximate that of the site into which the solid
substrate will be implanted, and may be on an order of magnitude scaling from millimeters
to centimeters, as needed. Similarly, shapes of the solid substrates of the invention
may be any shape into which the solid substrates of this invention may be machined
or processed, and may have any configuration as will be appropriate to achieve the
desired growth, repair or regeneration of bone and/or cartilage.
[0104] The invention provides a kit for the repair of cartilage comprising the solid substrates
of this invention, directions for utilizing said solid substrate in the repair of
cartilage and optionally a tool or tools for optimal insertion of said solid substrate,
seeding said solid substrate with cells or a combination thereof.
[0105] The kit of this invention comprises:
- a solid substrate comprising:
❖ a first phase comprising a marine organism skeletal derivative -based solid substrate
comprising a series of hollows along a longitudinal axis in said first phase and a
second biocompatible polymer located within the series of hollows along a longitudinal
axis; and
❖ a second phase comprising a marine organism skeletal derivative -based solid substrate;
❖ wherein said marine organism skeletal derivative-based solid substrate is a coral
or coral derivative, wherein said coral or coral derivative is aragonite, calcite,
hydroxyapatite, mixtures thereof, or other polymorphs of the same, and
- at least one first biocompatible polymer which is an elastic material which is less
rigid in structure than that of a first and a second phase
and wherein the kit is provided with instructions and appropriate tools for applying
the second biocompatible polymer to the solid substrate such that some of the second
biocompatible polymer is substantially located within the series of hollows in the
substrate and such that some of the first biocompatible polymer creates a separate
phase at a terminus of the solid substrate and the first biocompatible polymer is
added to said solid substrate in situ, and wherein said marine organism skeletal derivative
is characterized by a specific fluid uptake capacity value of at least 75%, which
specific fluid uptake capacity value is determined by establishing a spontaneous fluid
uptake value divided by a total fluid uptake value, wherein said fluid is a protein-containing,
salt-containing or carbohydrate containing solution, or wherein said fluid is a hydrophilic
biologic fluid, wherein said biologic fluid is autologous with respect to a cell or
tissue of a subject when said solid substrate is contacted with a cell or tissue of
said subject or wherein said fluid is water, or wherein said marine organism skeletal
derivative is characterized by having a contact angle value of less than 60 degrees,
when in contact with a fluid, wherein said fluid is a protein-containing, salt-containing
or carbohydrate containing solution, or wherein said fluid is a hydrophilic biologic
fluid, wherein said biologic fluid comprises cells or tissue, or wherein said fluid
is water.
[0106] According to the invention, such kits are provided with instructions and appropriate
tools for applying the second biocompatible polymer to the solid substrate, such that
some of the second biocompatible polymer is substantially located within the series
of hollows in the substrate and such that some of the first biocompatible polymer
creates a separate phase at a terminus of such solid substrate.
[0107] In one embodiment, the coral is seeded with a precursor cell. In one embodiment,
the precursor cell is a mesenchymal stem cell. In other embodiments, the cell may
be a mesenchymal cell; chondrocyte; fibrochondrocyte; osteocyte; osteoblast; osteoclast;
synoviocyte; bone marrow cell; stromal cell; stem cell; embryonic stem cell; precursor
cell, derived from adipose tissue; peripheral blood progenitor cell; stem cell isolated
from adult tissue; genetically transformed cell; or a combination thereof. In another
embodiment, a precursor cell may refer to a combination of chondrocytes and other
cells; a combination of osteocytes and other cells; a combination of synoviocytes
and other cells; a combination of bone marrow cells and other cells; a combination
of mesenchymal cells and other cells; a combination of stromal cells and other cells;
a combination of stem cells and other cells; a combination of embryonic stem cells
and other cells; a combination of precursor cells isolated from adult tissue and other
cells; a combination of peripheral blood progenitor cells and other cells; a combination
of stem cells isolated from adult tissue and other cells; and a combination of genetically
transformed cells and other cells. In one embodiment of the present invention, the
precursor cells for use in the method of the present invention are prepared from an
organ tissue of the recipient mammal (i.e. autologous), or a syngeneic mammal. In
another embodiment, allogeneic and xenogeneic precursor cells may be utilized.
[0108] In one embodiment, a site of cartilage repair may be considered to comprise a 3 dimensional
(3-D) space at or proximal to a site of a cartilage and/or defect or potential defect.
In one embodiment, this 3-D space comprises at least a wall or a floor, or a combination
thereof, and positioning within such a site may be described herein, relative to said
wall or floor, or in some embodiments, positioning may be relative to insertion within
a tissue site proximal to said wall or floor. In some embodiments, positioning include
insertion of the solid substrate or a region thereof, past the wall and/ or floor
of cartilage and/or bone tissue or a site of defect or injury or potential defect
or injury in the cartilage and/or bone tissue, such that insertion into bone tissue
occurs.
[0109] One skilled in the art will recognize that the shape of a site of cartilage and/or
bone repair and the shape of a 3-D solid substrate of this invention provide many
different combinations for stably positioning a solid substrate within a site of cartilage
repair and/or bone. In one embodiment, a solid substrate of this invention is shaped
prior to use in methods of this invention for cartilage repair and/or bone. In one
embodiment, a solid substrate of this invention is shaped concurrent to use in methods
of this invention for cartilage and/or bone repair. By shaping a solid substrate concurrent
with use of the solid substrate in methods of this invention, the dimensions of the
solid substrate may be precisely selected for specific positioning of the solid substrate
within a site of repair. It will be appreciated that multiple solid substrates of
this invention may be placed within or shaped and placed within a site of cartilage
and/or bone repair.
[0110] In some embodiments, reference to a "solid substrate", "implant" or "plug", as used
herein refers to any embodiment or combined embodiments as herein described with regard
to the solid substrates to be considered as being included in the described aspect
of this invention. For example, reference to a "solid substrate" as used herein, is
to be understood to refer to any embodiment of a solid substrate as described herein
being applicable for the indicated purpose or containing the indicated attribute,
etc.
[0111] In one embodiment, "solid substrate" refers to a shaped platform used for cartilage
and/or bone repair, wherein the shaped platform provides a site for cartilage and/or
bone regeneration. In one embodiment, the solid substrate is a temporary platform.
In one embodiment, "temporary platform" refers to a natural degradation of a coral
of this invention that occurs over time during cartilage and/or bone repair, wherein
the natural fully or partially degradation of the coral may results in a change of
solid substrate shape over time and/or change in solid substrate size over time.
[0112] In one embodiment, the coral is shaped in the form of the tissue to be grown. For
example, the coral can be shaped as a piece of cartilaginous tissue, such as a meniscus
for a knee or elbow; a joint; an articular surface of a bone, the rib cage, a hip,
a pelvis, an ear, a nose, a ligament, the bronchial tubes and the intervertebral discs.
[0113] This invention provides, in some embodiments, coral solid substrates for use in repairing
cartilage and/or bone tissue defects associated with physical trauma, or cartilage
and/or bone tissue defects associated with a disease or disorder in a subject.
[0114] In one embodiment of this invention, the term "coral" refers to coral which is cut
from a single piece of coral. In one embodiment, the coral has pore-like cavities
or interstices.
[0115] In one embodiment, the coral solid substrate is shaped prior to use in a method of
cartilage and/or bone repair. In one embodiment, the coral solid substrate is shaped
concurrent with a method of cartilage and/or bone repair, e.g., the coral solid substrate
may be shaped during surgery when the site of repair may be best observed, thus optimizing
the shape of the solid substrate used.
[0116] The solid substrates and/or kits of this invention employ use of a coral. In one
embodiment, the coral comprise any species, including, inter alia, Porites, Acropora,
Goniopora, Millepora, or a combination thereof.
[0117] In one embodiment, the coral is from the Porites species. In one embodiment, the
coral is
Porites Lutea. In most species, the void to solid ratios is generally in the range of 0.4 to 0.6,
and the void phase completely interconnects, forming a highly regular network that
interpenetrates the solid calcium carbonate phase. In one embodiment, this uniform
and interconnecting architecture is particularly useful as a framework in the solid
substrates, methods and/or kits of this invention.
[0118] In one embodiment, the coral is from the Goniopora species. In some embodiments,
the coral is
Goniopora albiconus, Goniopora burgosi, Goniopora cellulosa, Goniopora ceylon, Goniopora
ciliatus, Goniopora columna, Goniopora djiboutiensis, Goniopora eclipsensis, Goniopora
fruticosa, Goniopora gracilis, Goniopora klunzingeri, Goniopora lobata,
Goniopora mauritiensis, Goniopora minor, Goniopora norfolkensis, Goniopora palmensis,
Goniopora pandoraensis, Goniopora parvistella, Goniopora pearsoni, Goniopora pendulus,
Goniopora planulata, Goniopora polyformis, Goniopora reptans, Goniopora savignyi,
Goniopora somaliensis, Goniopora stokes, Goniopora stutchburyi, Goniopora sultani,
Goniopora tenella, Goniopora tenuidens or Goniopora viridis.
[0119] In another embodiment, the coral is from any one or more of the following species
Favites halicora; Goniastrea retiformis; Acanthastrea echinata; Acanthastrea hemprichi;
Acanthastrea ishigakiensis; Acropora aspera; Acropora austera; Acropora sp. "brown
digitate"; Acropora carduus; Acropora cerealis; Acropora chesterfieldensis; Acropora
clathrata; Acropora cophodactyla; Acropora sp. "danai-like"; Acropora divaricata;
Acropora donei; Acropora echinata; Acropora efflorescens; Acropora gemmifera; Acropora
globiceps; Acropora granulosa; Acropora cf hemprichi; Acropora kosurini; Acropora
cf loisettae; Acropora longicyathus; Acropora loripes; Acropora cf lutkeni; Acropora
paniculata; Acropora proximalis; Acropora rudis; Acropora selago; Acropora solitaryensis;
Acropora cf spicifera as per Veron; Acropora cf spicifera as per Wallace; Acropora
tenuis; Acropora valenciennesi; Acropora vaughani; Acropora vermiculata; Astreopora
gracilis; Astreopora myriophthalma; Astreopora randalli ; Astreopora suggesta; Australomussa
rowleyensis; Coscinaraea collumna; Coscinaraea crassa; Cynarina lacrymalis; Distichopora
violacea; Echinophyllia echinata; Echinophyllia cf echinoporoides; Echinopora gemmacea;
Echinopora hirsutissima; Euphyllia ancora; Euphyllia divisa; Euphyllia yaeyamensis;
Favia rotundata; Favia truncatus; Favites acuticollis; Favities pentagona; Fungia
granulosa; Fungia klunzingeri; Fungia mollucensis; Galaxea acrhelia; Goniastrea edwardsi;
Goniastea minuta; Hydnophora pilosa; Leptoseris explanata; Leptoseris incrustans;
Leptoseris mycetoseroides; Leptoseris scabra; Leptoseris yabei; Lithophyllon undulatum;
Lobophyllia hemprichii; Merulina scabricula; Millepora dichotoma; Millepora exaesa;
Millipora intricata; Millepora murrayensis; Millipora platyphylla; Monastrea curta;
Monastrea colemani; Montipora caliculata; Montipora capitata; Montipora foveolata;
Montipora meandrina; Montipora tuberculosa; Montipora cf vietnamensis; Oulophyllia
laevis; Oxypora crassispinosa; Oxypora lacera; Pavona bipartita; Pavona venosa; Pectinia
alcicornis; Pectinia paeonea; Platygyra acuta; Platygyra pini; Platygyra sp "green";
Platygyra verweyi; Podabacia cf lanakensis; Porites annae; Porites cylindrica; Porites
evermanni; Porites monticulosa; Psammocora digitata; Psammocora explanulata; Psammocora
haimeana; Psammocora superficialis; Sandalolitha dentata; Seriatopora caliendrum;
Stylocoeniella armata; Stylocoeniella guentheri; Stylaster sp.; Tubipora musica; Turbinaria
stellulata; or any coral known in the art, or a combination thereof.
[0120] In another disclosure, derivatives of marine animals such as coral, sponges, moluscs
shells and other related organisms may be used in the solid substrates, methods and/or
kits of this invention. Such derivatives may comprise,
inter alia, derivatives of Madreporaria, Helioporida of the order Coenothecalia, Tubipora of
the order Stolonifera, Millepora of the order Milleporina, or others known in the
art. In some embodiments, coral for use in the substrates, methods and/or kits of
this invention may comprise scleractinian coral, including in some embodiments, Goniopora
and others. In some embodiments, coral for use in the substrates, methods and/or kits
of this invention may comprise Alveoppora. In some embodiments, coral for use in the
substrates, methods and/or kits of this invention may comprise bamboo corals, including
in some embodiments, coral from the family Isididae, genera Keratoisis, Isidella,
and others.
[0121] In one embodiment of this invention, the term "coral" refers to coral which is cut
from a single piece of coral.
[0122] In some embodiments, the solid substrate is of any desired shape.
[0123] In one embodiment, coral may be machined into a variety of configurations, and quite
complex shapes such as cylindrical structures and threaded structures may be formed
by appropriate machine or other processing, such as chemical processing. In another
embodiment, coral may be shaped to form solid blocks, rods or granular forms. In one
embodiment, coralline materials are shaped in such a way as to conform to the shape
of a desired tissue structure or to fill gap and contour defects in a potential implantation
site. In one embodiment, coral is implanted in an orientation that allows it to contact
the maximum surface area of an adjacent-located tissue structure.
[0124] As described above, a solid substrate's region's ability to position and confine
the solid substrate of this invention is dependent on the region's geometry and the
geometry at the site of cartilage and/or bone repair where the solid substrate will
be implanted. In one embodiment, the region's geometry comprises a sharp edge. In
one embodiment, the region's geometry comprises a rounded edge. In one embodiment,
the region's geometry comprises a jagged edge.
[0125] In one disclosure of this invention, an optimal depth and angle within a site of
cartilage and/or bone repair comprise the depth and angle most beneficial for cartilage
and/or bone repair. In one disclosure, the optimal depth and angle most beneficial
comprise a position so that a solid substrate of this invention is accessible to a
pool of mesenchymal stem cells, a tissue milieu, blood vessels, nutrients, an effector
compound, or a therapeutic compound, or a combination thereof.
[0126] In one disclosure of this invention, the term "depth" refers to a measurement of
a solid substrate of this invention extending from an imaginary line resting on the
open surface of a repair site to a place beneath the tissue floor at a site of cartilage
and/or bone repair.
[0127] It will be recognized by one skilled in the art that the depth of other regions of
the solid substrate may not be below any tissue surface. For example based on a site
of cartilage repair shaped like a cylindrical pit, an imaginary line drawn to rest
across the opening of the pit represents the top of the pit. In one embodiment, positioning
of the solid substrate results in the entirety of the solid substrate being below
the top of the pit and therefore at a depth below the imaginary line across the opening.
Positioning of the solid substrate results in a portion of the solid substrate being
flush with the top of the pit.
[0128] In some disclosures, multiple solid substrates are inserted to maximally occupy a
defect site, such that each solid substrate material may be inserted at a different
angle and/or shape and/or depth and/or porosity to accommodate proper insertion into
the desired region within a site of cartilage and/or bone repair. It is to be understood
that the reference to angles of positioning above may be with regard to one or more
solid substrates inserted in a particular cartilage and/or bone defect site.
[0129] Contact between exposed surfaces of a solid substrate and tissue at or proximal to
a site of cartilage and/or bone repair provides a bioactive surface which, in the
methods of use of this invention may induce or enhance cartilage and/or bone repair.
For example the exposed surface of a solid substrate provides a bioactive surface
attracting mesenchymal stem cells. In another embodiment, the exposed surface provides
a place for mesenchymal stem cell attachment, growth, proliferation, or differentiation,
or a combination thereof, all processes which induce or enhance cartilage repair.
In addition, the exposed surface of a solid substrate may attract blood vessels. Moreover,
tissue at or proximal to a site of cartilage and/or bone repair may be a rich source
of nutrients, effector compounds, therapeutic compounds, or a combination thereof,
which may be beneficial in cartilage and/or bone repair so that contact between an
exposed surface of a solid substrate and such tissue induces or enhances cartilage
and/or bone repair.
[0130] Thus, it will be apparent to one skilled in the art that the specific positioning
of the solid substrate within a site of cartilage and/or bone repair arranges the
solid substrate of this invention such that the solid substrate is most effective
for cartilage and/or bone repair.
[0131] In one embodiment, "solid substrate" refers to a shaped platform used for cartilage
and/or bone repair, wherein the shaped platform provides a site for cartilage and/or
bone formation and/or regeneration. In one embodiment, the solid substrate is a temporary
platform. In one embodiment, "temporary platform" refers to a natural fully or partially
degradation of a coral of this invention that occurs over time during cartilage repair,
wherein the natural degradation of the coral may results in a change of solid substrate
shape over time and/or a change in solid substrate size over time.
[0132] In one embodiment, the coral is shaped in the form of the tissue to be grown. For
example, the coral can be shaped as a piece of cartilaginous or bony tissue, such
as a meniscus for a knee or elbow; a joint; an articular surface of a bone, the rib
cage, a hip, a pelvis, an ear, a nose, the bronchial tubes, the intervertebral discs,
a ligament, a vertebra, the tibia, the femur, the shoulder and the jaw.
[0133] In some disclosures, coral solid substrates for use in repairing cartilage and/or
bone tissue defects associated with physical trauma, or cartilage and/or bone tissue
defects associated with a disease or disorder in a subject.
[0134] In one embodiment of this invention, the term "coral" refers to coral which is cut
from a single piece of coral. In one embodiment, the coral has pore-like cavities
or interstices.
[0135] In one disclosure, the coral solid substrate is shaped prior to use in a method of
cartilage and/or bone repair. In one disclosure, the coral solid substrate is shaped
concurrent with a method of cartilage and/or bone repair, e.g., the coral solid substrate
may be shaped during surgery when the site of repair may be best observed, thus optimizing
the shape of the solid substrate used.
[0136] In one embodiment, the size of a solid substrate may be any size that would be useful
for the purposes of the present invention, as would be known to one skilled in the
art. In one embodiment, the solid substrate or a portion thereof may be about the
size of a site of cartilage and/or bone repair. In one embodiment, the solid substrate
or a portion thereof may be about the size of a cartilage and/or bone defect so that
the solid substrate may be placed within a site of cartilage and/or bone repair. In
another embodiment, the solid substrate may be larger than the size of a cartilage
and/or bone defect. For example, in one embodiment, the solid substrate of this invention
may be larger than the size of a cartilage and/or bone defect, whereby the solid substrate
may extend to a site of mesenchymal cell availability. In one embodiment, the solid
substrate may be smaller than the size of a cartilage and/or bone defect.
[0137] In some embodiments, the total solid substrate size will be on a millimeter scale,
for example, having at least one long axis of about 2-200 mm, or in some embodiments,
about 1-18 mm, or in some embodiments, about 0.5mm- 3 mm, or in some embodiments,
about 6-12 mm, or in some embodiments, about 10-15 mm, or in some embodiments, about
12-40 mm, or in some embodiments, about 30-100 mm, or in some embodiments, about 50-150
mm, or in some embodiments, about 100-200 mm.
[0138] In some embodiments the total solid substrate size will be on the centimeter scale,
for example having at least one long axis of about 0.5-30 cm
[0139] In one embodiment, the solid substrate may be about the same size as a tissue void
at a site of tissue repair. This tissue void may be due to a cartilage and/or bone
defect, cartilage and/or bone degeneration or may have been created artificially during
methods of cartilage and/or bone repair or any combination thereof. In one embodiment,
the tissue void comprises an absence of cartilage and/or bone tissue. In one embodiment,
the solid substrate or a portion thereof may be the size of a cartilage and/or bone
defect such that the solid substrate may be placed within a site of cartilage and/or
bone repair to enhance cartilage and/or bone formation at the site of cartilage and/
or bone repair. In another embodiment, the solid substrate may be larger than the
size of a cartilage and/or bone defect so that the solid substrate may reach to a
site of mesenchymal stem cell availability.
[0140] In some embodiments, a tight fit is desirable with regard to fitting the implant
within the site of tissue repair, except that it is desirable that the solid substrate
be below the upper void limit or flush with the same. According to this aspect, and
in some embodiments, it may be desirable to taper a terminus of the solid substrates
of this invention for easy insertion within a tight space for optimal tight fitting
of the implant.
[0141] In one embodiment, the term "void" refers to a space not occupied. In the instant
invention, for example, in one embodiment, a void may be a space in a solid substrate
naturally not occupied. In one embodiment, a void may be a space not occupied at a
site of repair. In one embodiment, a void may be a space not occupied within a solid
substrate of the current invention. In one embodiment, a void may be a volume of a
pore or a pore region.
[0142] In one embodiment, coral is washed, bleached, frozen, dried, sterilized or a combination
thereof. In some embodiments, the coral is processed as exemplified further hereinunder.
In some embodiments, the coral, once processed into the solid substrates of this invention
are seeded with a desired population of cells or populations of cells, prior to implantation
within a site of cartilage and/or bone repair.
[0143] In one embodiment, the multi-phasic solid substrate for the repair of cartilage is
prepared by an appropriate process known such as for example, the process described
in
WO 2010/146575.
[0144] In one embodiment of this invention, the term "portion" refers to a limited part
of a whole. In one embodiment, the term "portion" with regard to the surface exposed
as a consequence of the methods of this disclosure refers to a limited part of a whole
exposed surface. In one embodiment of this invention, the term "surface" refers to
an exterior or upper boundary of an object.
[0145] In one embodiment of this invention, the term "exposed" refers to being open to the
surrounding environment such that contact may occur between a solid substrate of this
invention and the immersion media.
[0146] According to this aspect, and in other embodiments, the solid forms of this invention
comprise multiple phases which phases differ in their pore volume, or which phases
comprise voids which differ in terms of the average diameter of said voids, or a combination
thereof.
[0147] In one embodiment, a solid substrate of this invention comprises a solid throughout
a solid substrate. One skilled in the art will recognize that solid solid substrateing
of this invention still comprises pore-like cavities and/or interstices.
[0148] A solid substrate of this invention comprises a hollow along a Cartesian coordinate
axis of a solid substrate. The hollow is along a long axis of a solid substrate of
this invention. The term "hollow" refers to a cavity within a solid substrate of this
invention. In one embodiment, the hollow comprises at least a single opening in the
solid substrate such that the cavity is exposed to the external environment. In one
embodiment, the hollow provides additional exposed surface area for a solid substrate
of this invention.
[0149] The solid substrates of this invention will comprise multiple hollows, which may
be in any orientation, or in some embodiments, the solid substrates of this invention
will comprise a network of hollows within solid substrates.
[0150] It will be appreciated by the skilled artisan that methods for selective creation
of hollows or voids (which words may be used interchangeably throughout) within the
solid substrates of this invention may be prepared by any means known to the skilled
artisan, for example, in accordance with the methods as herein described, for example,
by replacing immersion dipping of the portion of the solid substrate with drip application
of the immersion solution to selectively create voids within the solid substrates
of this application.
[0151] The exposed surface area of a solid substrate of this invention provides a location
for mesenchymal stem cells, chondrocytes, osteoblasts, etc., attachment, growth, proliferation
or differentiation, or a combination and a location for blood vessels formation. Therefore,
the surface area of a solid substrate of this invention ultimately provides a beneficial
location for regeneration of cartilage and/or bone tissue. In one embodiment of this
invention, a solid substrate comprises a hollow, wherein the presence of the hollow
increases the exposed surface area of a solid substrate compared to an analogous solid
substrate without a hollow.
[0152] According to this invention, the solid substrate comprises biocompatible polymers.
[0153] The term "biocompatible polymers" refers, in some embodiments, to the presence of
a polymeric material in association with at least a portion of the scaffolding material,
existing as part of the discrete phases as herein described.
[0154] In some embodiments, the solid substrate incorporates a biocompatible polymer therewithin,
which is associated with the aragonite or calcite component, via any physical or chemical
association. In some embodiments, the polymer is a part of a hydrogel, which is incorporated
in the solid substrates of this invention. In some embodiments, such hydrogel-containing
solid substrates may thereafter be lyophilized or dessicated, and may thereafter be
reconstituted.
[0155] In one embodiment, the biocompatible polymers of this invention comprise a natural
polymer comprising, collagen, elastin, silk, hyaluronic acid, sodium hyaluronate,
cross linked hyalronic acid, chitosan, cross linked chitosan, alginate, calcium alginate,
cross linked calcium alginate and any combinations thereof.
[0156] In one embodiment of this invention, a polymer comprises proteins such as zein, modified
zein, casein, gelatin, gluten, serum albumin, collagen, actin, α-fetoprotein, globulin,
macroglobulin, cohesin, laminin, fibronectin, fibrinogen, osteocalcin, osteopontin,
osteoprotegerin, or others, as will be appreciated by one skilled in the art. In another
embodiment, a polymer may comprise cyclic sugars, cyclodextrins, synthetic derivatives
of cyclodextrins, glycolipids, glycosaminoglycans, oligosaccharide, polysaccharides
such as alginate, carrageenan (χ, λ, µ, κ), chitosane, celluloses, condroitin sulfate,
curdlan, dextrans, elsinan, furcellran, galactomannan, gellan, glycogen, arabic gum,
hemicellulose, inulin, karaya gum, levan, pectin, pollulan, pullulane, prophyran,
scleroglucan, starch, tragacanth gum, welan, xanthan, xylan, xyloglucan, hyaluronic
acid, chitin, or a poly(3-hydroxyalkanoate)s, such as poly(P-hydroxybutyrate), poly(3-hydroxyoctanoate)
or poly(3-hydroxyfatty acids), or any combination thereof.
[0157] In one embodiment, a polymer may comprise chemical derivatives thereof (substitutions,
additions, and elimination of chemical groups, for example, alkyl, alkylene, hydroxylations,
oxidations, and other modifications routinely made by those skilled in the art), blends
of, e.g. proteins or carbohydrates alone or in combination with synthetic polymers.
[0158] In one embodiment of this invention, the polymer is biodegradable. In one embodiment,
the term "biodegradable" or grammatical forms thereof, refers to a material of this
invention, which is degraded in the biological environment of the subject in which
it is found. In one embodiment, the biodegradable material undergoes degradation,
during which, acidic products, or in another embodiment, basic products are released.
In one embodiment, bio-degradation involves the degradation of a material into its
component subunits, via, for example, digestion, by a biochemical process. In one
embodiment, biodegradation may involve cleavage of bonds (whether covalent or otherwise),
for example in a polymer backbone of this invention. In another embodiment, biodegradation
may involve cleavage of a bond (whether covalent or otherwise) internal to a side-chain
or one that connects a side chain to, for example a polymer backbone.
[0159] In one embodiment, a coral of this invention is covalently associated with the polymer
via the use of a cross-linking agent. In one embodiment, the phrase "cross-linking
agent" refers to an agent which facilitates the formation of a covalent bond between
2 atoms. In one embodiment, the cross-linking agent is a zero-length cross-linking
agent.
[0160] In one embodiment, the cross-linking agent is (1 ethyl 3-(3dimethyl aminopropyl)carbodiimide
(EDAC), N-Sulfohydroxy succinamide (Sulfo NHS), 5-iodopyrimidines, N-carbalkoxydihydroquinolines,
pyrroloquinolinequinones, or a combination thereof.
[0161] In one embodiment, the cross-linking agent is a homobifunctional cross-linker, such
as, for example, a N-hydroxysuccinimide ester (e.g. disuccinimidyl suberate or dithiobis(succinimidylpropionate),
homobifunctional imidoester (e.g. dimethyladipimidate or dimethyl pimelimidate), sulfhydryl-reactive
crosslinker (e.g. 1,4-di-[3'-(2'-pyridyldithio)propionamido]butane), difluorobenzene
derivative (e.g.1,5-difluoro-2,4-dinitrobenzene), aldehyde (e.g. formaldehyde, glutaraldehyde),
bis-epoxide (e.g. 1,4-butanediol diglycidyl ether), hydrazide (e.g. adipic acid dihydrazide),
bis-diazonium derivative (e.g. o-tolidine), bis-alkylhalide, or a combination thererof.
[0162] In one embodiment, the cross-linking agent is a heterobifunctional cross-linker,
such as, for example, an amine-reactive and sulfhydryl-reactive crosslinker (e.g.
N-succinimidyl 3-(2-pyridyldithio)propionate, a carbonyl-reactive and sulfhydryl-reactive
crosslinker (e.g. 4-(4-N-maleimidophenyl)butyric acid hydrazide), or a combination
thereof.
[0163] In some embodiments, the cross-linking agent is a trifunctional cross-linkers, such
as, for example, 4-azido-2-nitrophenylbiocytin-4-nitrophenyl ester, sulfosuccinimidyl-2-[6-biotinamido]-2-(p-azidobenzamido)hexanoamido]ethyl-1,3'-dithiopropionate
(sulfo-SBED), or a combination thereof.
[0164] In another embodiment, the cross-linking agent is an enzyme. In one embodiment of
this invention, the cross-linking agent comprises a transglutaminase, a peroxidase,
a xanthine oxidase, a polymerase, or a ligase, or a combination thereof.
[0165] The choice of concentration of the cross-linking agent utilized for activity will
vary, as a function of the volume, agent and polymer chosen, in a given application,
as will be appreciated by one skilled in the art.
[0166] In one embodiment, the association of a coral of this invention with a biocompatible
polymer of this invention comprises a physical and/or mechanical association. For
example, in one embodiment, a physical and/or mechanical association may comprise
imbibing of any means, air drying, using a cross-linking agent, applying of heat,
applying vacuum, applying lyophilizing methods, freezing, applying mechanical forces
or any combination thereof, to promote the physical association between a coral and
a polymer coating as described herein.
[0167] In one embodiment, the thickness of the first phase containing the biocompatible
polymer influences physical characteristics of a solid substrate of this invention.
For example, the thickness of the polymer phase may influence elasticity, tensile
strength, adhesiveness, or retentiveness, or any combination thereof of a solid substrate
of this invention. In one embodiment, the thickness of the first phase containing
the biocompatible polymer is selected so that it increases the elasticity of a solid
substrate of this invention. In one embodiment, the thickness of the first phase containing
the biocompatible polymer is selected so that it increases the tensile strength of
a solid substrate of this invention. In one embodiment, the thickness of the first
phase containing the biocompatible polymer is selected so that it affects adhesion
of mesencymal stem cells, blood vessels, tissue at a site of cartilage repair, cartilage
tissue, or bone tissue, or a combination thereof. In one embodiment, the thickness
of the first phase containing the biocompatible polymer is selected so that it decreases
the adhesiveness of a solid substrate of this invention. In one embodiment, the thickness
of the first phase containing the biocompatible polymer is selected so that it increases
the adhesiveness of a solid substrate of this invention.
[0168] In one embodiment, the thickness of the first phase containing the biocompatible
polymer is selected so that it influences proliferation and/or differentiation of
mesenchymal stem cells applied to the solid substrates of this invention, or influences
the activation or migration of cells associated with cartilage and/or bone formation
or repair to the solid substrates of this invention, or a combination thereof.
[0169] In one embodiment of this invention, the cells as used in accordance with the solid
substrates or kits of this invention, are engineered to express a desired product.
[0170] In one embodiment, the biocompatible polymers of this invention comprise an effector
compound. In one embodiment, the effector compound comprises a component of a kit
of this invention for use for incorporation into a solid substrate of this invention
as herein described. In one embodiment, the effector compound is applied directly
to a polymer coating of this invention, without being dispersed in any solvent.
[0171] In one embodiment of this invention, the biocompatible polymers of this invention
comprise an effector compound comprising a cytokine, a bone morphogenetic protein
(BMP), growth factors, a chelator, a cell population, a therapeutic compound, or an
antibiotic, or any combination thereof.
[0172] In one embodiment, effector compounds for use in a solid substrate and/or a kit of
this invention and/or a method of this invention may comprise,
inter-alia, a cytokine, a bone morphogenetic protein (BMP), growth factor, a chelator, a cell
population, a therapeutic compound, an anti-inflammatory compound, a pro-angiogenic
compound or an antibiotic, or any combination thereof.
[0173] In one embodiment, the phrase "a cell population" refers to a transfected cell population,
a transduced cell population, a transformed cell population, or a cell population
isolated from a subject, or a combination thereof. In some embodiments, transfected,
transduced or transformed cells, may be incorporated into a polymer coat, or a solid
substrate of this invention, or a combination thereof.
[0174] In one embodiment, transfected, transduced or transformed cells, may be incorporated
into a polymer coating, or a solid substrate of this invention
[0175] In one embodiment, a cell population of this invention comprises mesenchymal stem
cells. In one embodiment, the mesenchymal stem cells are transformed. In one embodiment,
a cell population comprises cells beneficial in cartilage and/or bone formation and/or
repair, such as chondroblasts or chondrocytes; fibrochondrocyte; osteocyte; osteoblast;
osteoclast; synoviocyte; bone marrow cell; stromal cell; stem cell; embryonic stem
cell; precursor cell, derived from adipose tissue; peripheral blood progenitor cell;
stem cell isolated from adult tissue; genetically transformed cell; or a combination
thereof. In another embodiment, a precursor cell may refer to a combination of chondrocytes
and other cells; a combination of osteocytes and other cells; a combination of synoviocytes
and other cells; a combination of bone marrow cells and other cells; a combination
of mesenchymal cells and other cells; a combination of stromal cells and other cells;
a combination of stem cells and other cells; a combination of embryonic stem cells
and other cells; a combination of precursor cells isolated from adult tissue and other
cells; a combination of peripheral blood progenitor cells and other cells; a combination
of stem cells isolated from adult tissue and other cells; and a combination of genetically
transformed cells and other cells. the precursor cells for use in the method of the
present invention are prepared from an organ tissue of the recipient mammal (i.e.
autologous), or a syngeneic mammal. In another embodiment, allogeneic and xenogeneic
precursor cells may be utilized.
[0176] In one embodiment of this invention, the phrase "a therapeutic compound" refers to
a peptide, a protein or a nucleic acid, or a combination thereof. In another embodiment,
the therapeutic compound is an antibacterial, antiviral, antifungal or antiparasitic
compound. In another embodiment, the therapeutic compound has cytotoxic or anti-cancer
activity. In another embodiment, the therapeutic compound is an enzyme, a receptor,
a channel protein, a hormone, a cytokine or a growth factor. In another embodiment,
the therapeutic compound is immunostimulatory. In another embodiment, the therapeutic
compound inhibits inflammatory or immune responses. In one embodiment, the therapeutic
compound comprises a pro-angiogenic factor.
[0177] In one embodiment, the phrase "a therapeutic compound", refers to a molecule, which
when provided to a subject in need, provides a beneficial effect. In some cases, the
molecule is therapeutic in that it functions to replace an absence or diminished presence
of such a molecule in a subject. In one embodiment, the molecule is a nucleic acid
coding for the expression of a protein is absent, such as in cases of an endogenous
null mutant being compensated for by expression of the foreign protein. In other embodiments,
the endogenous protein is mutated, and produces a non-functional protein, compensated
for by the expression of a heterologous functional protein. In other embodiments,
expression of a heterologous protein is additive to low endogenous levels, resulting
in cumulative enhanced expression of a given protein. In other embodiments, the molecule
stimulates a signaling cascade that provides for expression, or secretion, or others
of a critical element for cellular or host functioning.
[0178] In another embodiment, the therapeutic compound may be natural or non-natural insulins,
amylases, proteases, lipases, kinases, phosphatases, glycosyl transferases, trypsinogen,
chymotrypsinogen, carboxypeptidases, hormones, ribonucleases, deoxyribonucleases,
triacylglycerol lipase, phospholipase A2, elastases, amylases, blood clotting factors,
UDP glucuronyl transferases, ornithine transcarbamoylases, cytochrome p450 enzymes,
adenosine deaminases, serum thymic factors, thymic humoral factors, thymopoietins,
growth hormones, somatomedins, costimulatory factors, antibodies, colony stimulating
factors, erythropoietin, epidermal growth factors, hepatic erythropoietic factors
(hepatopoietin), liver-cell growth factors, interleukins, interferons, negative growth
factors, fibroblast growth factors, transforming growth factors of the α family, transforming
growth factors of the β family, gastrins, secretins, cholecystokinins, somatostatins,
serotonins, substance P, transcription factors or combinations thereof.
[0179] In one embodiment, the effector compound comprises, an anti-helminth, an antihistamine,
an immunomodulatory, an anticoagulant, a surfactant, an antibody, a beta-adrenergic
receptor inhibitor, a calcium channel blocker, an ace inhibitor, a growth factor,
a hormone, a DNA, an siRNA, or a vector or any combination thereof.
[0180] In one embodiment, the phrase "effector compound" refers to any agent or compound,
which has a specific purpose or application which is useful in the treatment, prevention,
inhibition, suppression, delay or reduction of incidence of infection, a disease,
a disorder, or a condition, when applied to the solid substrates, kits and/or methods
of this invention. An effector compound of this invention, in one embodiment, will
produce a desired effect which is exclusive to the ability to image the compound.
In some embodiments, the effector compound may be useful in imaging a site at which
the compound is present, however, such ability is secondary to the purpose or choice
of use of the compound.
[0181] In one embodiment of this invention, term "effector compound" is to be understood
to include the terms "drug" and "agent", as well, when referred to herein, and represents
a molecule whose incorporation within the solid substrate and/or kits of this invention,
or whose use thereof, is desired. In one embodiment, the agent is incorporated directly
within a solid substrate, and/or kit of this invention. In another embodiment, the
agent is incorporated within a solid substrate and/or kit of this invention, either
by physical interaction with a polymer coating, a coral, or coral particles of this
invention, and/or a kit of this invention, or association thereto.
[0182] In one embodiment, compounds for use in a solid substrate and/or a kit of this invention
and/or a method of this disclosure may comprise,
inter-alia, an antibody or antibody fragment, a peptide, an oligonucleotide, a ligand for a biological
target, an immunoconjugate, a chemomimetic functional group, a glycolipid, a labelling
agent, an enzyme, a metal ion chelate, an enzyme cofactor, a cytotoxic compound, a
bactericidal compound, a bacteriostatic compound, a fungicidal compound, a fungistatic
compound, a chemotherapeutic, a growth factor, a hormone, a cytokine, a toxin, a prodrug,
an antimetabolite, a microtubule inhibitor, a radioactive material, or a targeting
moiety, or any combination thereof.
[0183] In one embodiment, the solid substrates and/or kits of this invention and/or methods
of this disclosure comprise or make use of an oligonucleotide, a nucleic acid, or
a vector. In some embodiments, the term "oligonucleotide" is interchangeable with
the term "nucleic acid", and may refer to a molecule, which may include, but is not
limited to, prokaryotic sequences, eukaryotic mRNA, cDNA from eukaryotic mRNA, genomic
DNA sequences from eukaryotic (e.g., mammalian) DNA, and even synthetic DNA sequences.
The term also refers to sequences that include any of the known base analogs of DNA
and RNA.
[0184] The solid substrates and/or kits of this invention and/or methods of use of this
disclosure may comprise nucleic acids, in one embodiment, or in another embodiment,
the solid substrates and/or kits of this invention and/or methods of use of this disclosure
may include delivery of the same, as a part of a particular vector. In one embodiment,
polynucleotide segments encoding sequences of interest can be ligated into commercially
available expression vector systems suitable for transducing/transforming mammalian
cells and for directing the expression of recombinant products within the transduced
cells. It will be appreciated that such commercially available vector systems can
easily be modified via commonly used recombinant techniques in order to replace, duplicate
or mutate existing promoter or enhancer sequences and/or introduce any additional
polynucleotide sequences such as for example, sequences encoding additional selection
markers or sequences encoding reporter polypeptides.
[0185] In one embodiment, the solid substrate of this invention incorporates stem or progenitor
or precursor cells. Such cells can be obtained directly from a mammalian donor, e.g.,
a patient's own cells, from a culture of cells from a donor, or from established cell
culture lines. In some embodiments, the mammal is a mouse, rat, rabbit, guinea pig,
hamster, cow, pig, horse, goat, sheep, dog, cat, monkey, ape or a human. Cells of
the same species and/or of the same immunological profile can be obtained by biopsy,
either from the patient or a close relative. Using standard cell culture techniques
and conditions, the cells are then grown in culture until confluent and used when
needed. The cells may be cultured until a sufficient number of cells have been obtained
for a particular application.
[0186] In one embodiment, the solid substrate of this invention incorporates any cell which
may participate in cartilage and/or bone formation or repair. In some embodiments,
such cells represent autografts, in that cells are cultured
ex-vivo to seed the cells on the solid substrates of the invention, and such seeded solid
substrates are implanted into the subject.
[0187] In some embodiments, such cells may represent allografts or xenografts, which may
be incorporated within the solid substrates of this invention and implanted within
a site of repair.
[0188] In one embodiment, a coral of this invention comprises a cell population from
in vitro culture of the coral for a time period sufficient to seed the cells in the coral.
In one embodiment, the cell population is a mesenchymal stem cell population, chondrocyte;
fibrochondrocyte; osteocyte; osteoblast; osteoclast; synoviocyte; bone marrow cell;
stromal cell; stem cell; embryonic stem cell; precursor cell, derived from adipose
tissue; peripheral blood progenitor cell; stem cell isolated from adult tissue; genetically
transformed cell; or a combination thereof. In one embodiment, the mesenchymal stem
cells; chondrocyte; fibrochondrocyte; osteocyte; osteoblast; osteoclast; synoviocyte;
bone marrow cell; stromal cell; stem cell; embryonic stem cell; precursor cell, derived
from adipose tissue; peripheral blood progenitor cell; stem cell isolated from adult
tissue; genetically transformed cell; or a combination thereof seeded
in vitro are transformed. In one embodiment, the cell population comprises a cell population
beneficial for cartilage repair. In one embodiment, the culture comprises a chelator.
In one embodiment of this invention, the chelator in a culture comprises a calcium
chelator.
[0189] According to this invention the marine organism skeletal derivative is characterized
by a specific fluid uptake capacity value of at least 75%, which specific fluid uptake
capacity value is determined by establishing a spontaneous fluid uptake value divided
by a total fluid uptake value.
[0190] According to this aspect, a specific fluid uptake capacity value may be determined
by evaluating spontaneous uptake of a biologic fluid versus a total uptake capacity
for a given sample and arriving at the specific fluid uptake capacity level, whereby
if the value is over 75%, then such solid substrate will be used as described.
[0191] In some embodiments, the process for selection of the material for incorporation
within a solid substrate of this invention comprises isolating a sample of a coralline-based
solid material and establishing a specific fluid uptake capacity value of the material,
which specific fluid uptake capacity value is determined as described.
[0192] In some embodiments, the biologic fluid is blood, and in some embodiments, the biologic
fluid is water. In some embodiments, the biologic fluid is hydrophilic.
[0193] In some embodiments, the biologic fluid is autologous with respect to a cell or tissue
of a subject when said solid substrate is contacted with such cell or tissue of said
subject.
[0194] It will be understood that the biologic fluid may be any fluid which is biocompatible
and whose incorporation is appropriate within a solid substrate for the desired application.
[0195] In some embodiments, the process further comprises the step of contacting the material
with a fluid for from 2 - 15 minutes to promote spontaneous fluid uptake of said fluid
within said coralline-based solid material to arrive at said spontaneous fluid uptake
value. In some embodiments, the process may allow for the contacting of the material
with a fluid for from 0.5 - 15 minutes, or in some embodiments, from 0.5 - 5 minutes,
or in some embodiments, 10 - 60 minutes, or in some embodiments, from 60 to 90 minutes,
or in some embodiments, other intervals, to promote spontaneous fluid uptake. The
skilled artisan will appreciate that the amount of time for which the fluid is applied
to determine the spontaneous uptake may be extended or shortened as a function of
the dimensions and geometry of the sample substrate being assessed. In some embodiments,
when a larger sample is being assessed, the process further comprises the step of
contacting the material with a fluid for from 2 - 24 hours to promote spontaneous
fluid uptake of said fluid within said coralline-based solid material to arrive at
said spontaneous fluid uptake value
[0196] In some embodiments, the process further comprises the step of contacting said marine
organism skeletal derivative -based solid material with a fluid and applying negative
pressure to said coralline-based solid material to promote maximal uptake of said
fluid within said coralline-based solid material to arrive at said total fluid uptake
value. In some embodiments, application of positive pressure is via the application
of a vacuum to the substrate immersed in the fluid, promoting entry of the fluid therewithin.
[0197] In some embodiments, the process may further comprise the step of contacting said
coralline-based solid material with a fluid and applying positive pressure to said
coralline-based solid material to promote maximal uptake of said fluid within said
coralline-based solid material to arrive at said total fluid uptake value. According
to this aspect, and in some embodiments, care will be taken to ensure that the application
of pressure does not in any way compromise the structural integrity of the solid substrate.
[0198] In some embodiments, application of positive pressure is via any manual means, for
example, via the use of any applicator, syringe, etc., gravitational pressure, and
others, as will be appreciated by the skilled artisan. In some embodiments, application
of positive pressure is via forced osmosis, centrifugation and others. In some embodiments,
combinations of the described methods and others are envisioned.
[0199] In some embodiments, the marine organism skeletal derivative is characterized by
having a contact angle value of less than 60 degrees, when in contact with a fluid.
[0200] In some embodiments, the solid substrates characterized by a contact angle value
of less than 60 degrees is comparable to samples having a specific fluid uptake capacity
value of at least 75%,.
[0201] Methods for determining a contact angle are well known, and any appropriate method
can be used.
[0202] Such solid substrates may be converted to hydroxyapatite prior to or following such
assessment of specific fluid uptake value or contact value, as described.
[0203] According to this aspect, and in one embodiment, some or all phases of the solid
substrates as herein defined, such as, for example, coral samples or nacre or others
as herein described are assessed by selecting a small dry sample for use in the processes
as herein described, whose region of isolation from a larger block may be ascertained,
in order to provide information regarding the characteristics of the area in the block
from which additional samples may be isolated and then used.
[0204] In some aspects, the sample is dried under vacuum and/or heated or pressurized or
steam treated.
[0205] In some embodiments, for aspects relating to a specific fluid uptake capacity value,
such value is a function of change in weight in said coralline-based solid material.
[0206] According to this aspect and in some embodiments, the dry weight for each sample
is recorded and fluid as described herein is added an assay container.
[0207] According to this aspect and in some embodiments, at least 1:1 ratio of the size
of the sample in mm to the volume of fluid added in ml is applied to the container.
In some embodiments, the amount of fluid applied is in excess, as compared to the
sample size.
[0208] According to this aspect and in some embodiments, once the initial fluid uptake is
assessed, according to this aspect and in some embodiments, the solid substrate sample
is then brought into contact with the fluid and the weight of the solid substrate
sample is assessed. In other embodiments the specific gravity is assessed by gradient
centrifugation of by the Archimedean principle.
[0209] According to this aspect and in some embodiments, spontaneous fluid uptake is assessed
and a spontaneous fluid uptake value is established, based on the change in weight
of the sample.
[0210] According to this aspect and in some embodiments, the specific fluid uptake capacity
value is a function of change in fluid volume of applied fluid to said marine organism
skeletal derivative -based solid material. According to this aspect, spontaneous fluid
uptake is assessed and a spontaneous fluid uptake value is established based on the
complete uptake of the volume applied to the sample.
[0211] According to this aspect and in some embodiments, the process then further comprises
contacting a significantly increased amount of fluid with the sample and applying
pressure thereto to promote maximal fluid uptake to the total fluid uptake capacity
of the sample.
[0212] According to this aspect and in some embodiments, as noted, such pressure may be
either positive or negative pressure, and the application time is for a period of
time sufficient to ensure maximal uptake of the applied fluid into the marine organism
skeletal derivative sample.
[0213] According to this aspect and in some embodiments, such time may include an interval
of from 0.5 - 60 minutes, or in some embodiments, when a larger sample is being assessed,
such time may include an interval of from 2 - 24 hours to to arrive at said spontaneous
fluid uptake value. It will be appreciated that the time intervals recited herein
are applicable for any embodiment with regard thereto as described herein. The skilled
artisan will appreciate that the amount of time for which the fluid is applied to
determine the full capacity fluid uptake may be extended or shortened as a function
of the dimensions and geometry of the sample substrate being assessed.
[0214] According to these aspects, the total fluid uptake capacity is thus assessed and
the specific fluid uptake capacity value is then determined.
[0215] In some embodiments, the invention specifically contemplates solid substrates having
a specific fluid uptake capacity value exceeding the cutoff value of 75%, for the
sample to be noted optimized as a solid substrate for promoting cell or tissue growth.
It will be appreciated that the invention contemplates the stated cutoff value for
promoting a reasonable value that reduces the presence of appreciable false positives,
i.e. solid substrates that are not as optimal for the stated applications.
[0216] In some embodiments, the invention specifically contemplates solid substrates characterized
by one or more phases therein comprising a marine organism skeletal derivative having
a contact angle value of less than 60 degrees, when in contact with a fluid. It will
be appreciated that the invention contemplates the stated cutoff value for promoting
a reasonable value that reduces the presence of appreciable false positives, i.e.
solid substrates that are not as optimal for the stated applications.
[0217] In some disclosures, when the sample is utilized
in vivo in subsequent applications, in some aspects, the sample is first contacted with autologous
biological fluids or materials from the host prior to implantation into the same,
verifying the observed enhanced fluid uptake phenotype as herein described.
[0218] In other embodiments the substrate may be a mixture of several marine originated
materials or a mixture of bone and coral granules or cartilage and coral granules.
In some embodiments, the solid substrate may be a composite material comprised of
multiple samples of the marine organism skeletal derivatives as herein described.
[0219] In some embodiments, according to this aspect, the disclosure provides a method of
inducing or enhancing repair, regeneration or enhancement of formation of cartilage,
bone or a combination thereof, said method comprising implanting in a subject, a solid
substrate as herein described within a site in need of repair, regeneration or enhancement
of formation of cartilage, bone or a combination thereof.
[0220] In some embodiments, the disclosure provides a method of inducing or enhancing repair,
regeneration or enhancement of formation of cartilage, bone or a combination thereof,
said method comprising:
o implanting within a site in need of repair, regeneration or enhancement of formation
of cartilage, bone or a combination thereof in a subject, a solid substrate comprising:
- a first phase comprising marine organism skeletal derivative -based solid substrate
comprising a first biocompatible polymer and said first phase further comprises a
series of hollows along a longitudinal axis in said first phase, wherein said first
biocompatible polymer is substantially located within said series of hollows; and
- a second phase comprising marine organism skeletal derivative -based solid substrate,
optionally comprising a series of hollows along a longitudinal axis in said second
phase;
whereby said implanting places said solid substrate below an upper limit of said site
in need of repair, and
- applying a second biocompatible polymer to a region above said first phase to be slightly
below, to be flush with or to slightly exceed an upper limit of said site in need
of repair.
[0221] In one embodiment, the phrase "cartilage repair" refers to restoring a cartilage
defect to a more healthful state. In one embodiment, restoring cartilage results in
regeneration of cartilage tissue. In one embodiment, restoring cartilage results in
regeneration of a full or partial thickness articular cartilage defect. In one embodiment,
restoring cartilage results in complete or partial regeneration of cartilage tissue
at a site of cartilage repair. In one embodiment, cartilage repair may result in restoration/repair
of missing or defective bone tissue, wherein repair of a cartilage defect necessitates
removal of bone tissue at a site of cartilage repair. In one embodiment, restoring
cartilage results in regeneration of osteochondral defect. In one embodiment, cartilage
repair comprises restoring cartilage defects of joints (e.g. knee, elbow, hip, shoulder
joints), of ears, of a nose, or of a wind pipe.
[0222] In one embodiment, the phrase "bone repair" refers to restoring a bone defect to
a more healthful state. In one embodiment, restoring bone results in regeneration
of bone tissue. In one embodiment, restoring bone results in the filling in of any
fracture or void within a bone tissue. In one embodiment, restoring bone results in
complete or partial regeneration of bone tissue at a site of bone repair. In one embodiment,
bone repair may result in restoration/repair of missing or defective bone tissue.
In one embodiment, bone repair comprises restoring bone defects of any bone, as needed.
[0223] In some embodiments, the phrase "bone repair" refers to the treatment of a subject
with osteoporosis, Paget's disease, fibrous dysplasias, or osteodystrophies. In another
embodiment, the subject has bone and/or cartilage infirmity. In another embodiment,
the subject has other bone remodeling disorders include osteomalacia, rickets, rheumatoid
arthritis, achondroplasia, osteochodrytis, hyperparathyroidism, osteogenesis imperfecta,
congenital hypophosphatasia, fribromatous lesions, multiple myeloma, abnormal bone
turnover, osteolytic bone disease, periodontal disease, or a combination thereof.
In one embodiment, bone remodeling disorders include metabolic bone diseases which
are characterized by disturbances in the organic matrix, bone mineralization, bone
remodeling, endocrine, nutritional and other factors which regulate skeletal and mineral
homeostasis, or a combination thereof. Such disorders may be hereditary or acquired
and in one embodiment, are systemic and affect the entire skeletal system.
[0224] The solid substrates and kits of the invention may also be used to enhance bone and/or
cartilage formation in conditions where a bone and/or cartilage deficit is caused
by factors other than bone remodeling disorders. Such bone deficits include fractures,
bone trauma, conditions associated with post-traumatic bone surgery, post-prosthetic
joint surgery, post plastic bone surgery, bone chemotherapy, post dental surgery and
bone radiotherapy. Fractures include all types of microscopic and macroscopic fractures.
In one embodiment, some examples of fractures includes avulsion fracture, comminuted
fracture, transverse fracture, oblique fracture, spiral fracture, segmental fracture,
displaced fracture, impacted fracture, greenstick fracture, torus fracture, fatigue
fracture, intraarticular fracture (epiphyseal fracture), closed fracture (simple fracture),
open fracture (compound fracture) and occult fracture. In one embodiment, fractures
meant to be treated using the methods of the present invention are non-union fractures.
[0225] In one embodiment, the solid substrates and kits of the invention may also be used
to augment long bone fracture repair; generate bone in segmental defects; provide
a bone graft substitute for fractures; facilitate tumor reconstruction or spine fusion;
provide a local treatment (by injection) for weak or osteoporotic bone, such as in
osteoporosis of the hip, vertebrae, or wrist, or a combination thereof. In another
embodiment, the solid substrates and kits of the invention may also be used in a method
to accelerate the repair of fractured long bones; treat of delayed union or non-unions
of long bone fractures or pseudoarthrosis of spine fusions; induce new bone formation
in avascular necrosis of the hip or knee, or a combination thereof.
[0226] In one disclosure, a method of this disclosure comprises inducing and enhancing cartilage
and/or bone repair wherein implanting a solid substrate of this invention within a
site of cartilage and/or bone repair influences and improves cartilage and/or bone
repair.
[0227] In one disclosure, a method of this disclosure induces or enhances cartilage and/or
bone repair, wherein the solid substrate attracts a population of cells to the solid
substrate, thereby influencing or improving cartilage and/or bone repair.
[0228] The 3-D architecture and chemical composition of a solid substrate of this invention
are of great importance for specifically positioning and confining a solid substrate
within a site of cartilage and/or bone repair; for cellular recognition, adhesion,
proliferation and differentiation of cell populations which induce or enhance cartilage
and/or bone repair or a combination thereof.
[0229] In one embodiment, a solid substrate of this invention utilized in a method of this
disclosure comprises a seeded cell population prior to being implanted in a subject.
In one embodiment, a method of this invention induces or enhances cartilage and/or
bone repair, wherein implanting in a subject a solid substrate of this invention promotes
adhesion, proliferation or differentiation, or a combination thereof of transformed
mesenchymal stem cells. In one disclosure, a method of this disclosure induces or
enhances cartilage and/or bone repair, wherein implanting in a subject a solid substrate
of this invention promotes blood vessel formation.
[0230] In one embodiment, a solid substrate utilized in a method of the present disclosure
may be used to adsorb or bind, and deliver, other therapeutically active substances
which assist in the cartilage and/or bone repair or regeneration process, or which
have other desired therapeutic activity. Such substances include, by way of example,
known synthetic or semisynthetic antibiotics which may be introduced into the pore
cavities of the shaped product or structure, or a growth factor such as transforming
growth factor or one of the bone morphogenic proteins which can be used to assist
or promote bone growth.
[0231] In any of the embodiments herein, solid substrates for use in the methods of the
present disclosure may further comprise, or be implanted with, other compounds such
as, for example, antioxidants, growth factors, cytokines, antibiotics, anti-inflammatories,
immunosuppressors, preservative, pain medication, other therapeutics, and excipient
agents. In one embodiment, examples of growth factors that may be administered in
addition to the HMG-CoA reductase inhibitor include, but are not limited to, epidermal
growth factor (EGF), transforming growth factor-alpha (TGF-β), transforming growth
factor-beta (TGF-β), human endothelial cell growth factor (ECGF), granulocyte macrophage
colony stimulating factor (GM-CSF), bone morphogenetic protein (BMP), nerve growth
factor (NGF), vascular endothelial growth factor (VEGF), fibroblast growth factor
(FGF), insulin-like growth factor (IGF), cartilage derived morphogenetic protein (CDMP),
platelet derived growth factor (PDGF), or any combinations thereof. Examples of antibiotics
include antimicrobials and antibacterials.
[0232] In one disclosure, a method of this disclosure comprises implanting a solid substrate
of this invention in a subject afflicted with a cartilage and/or bone defect or disorder
or disease
[0233] In one embodiment, the term "implanting" refers to inserting and fixing a solid substrate
of this invention with in a living site in a subject, the site comprising a site of
cartilage and/or bone repair. In one disclosure, a method of this disclosure implants
a solid substrate such a region of the solid substrate now has access to mesenchymal
stem cells, nutrients, blood vessels, or effector compounds, or any combination there
of. In one disclosure, a method of this disclosure comprises implanting in a subject
a solid substrate of this invention, wherein the method results in removing a region
of cartilage and/or bone and/or other tissue so that a region of the solid substrate
penetrates through the cartilage and/or bone and/or other tissue, and in some disclosures,
reaches a bone marrow void.
[0234] A clinician skilled in the art will recognize that methods of this disclosure, which
entail implanting a solid substrate within a site of cartilage and/or bone repair,
may require preparation of a site of cartilage and/or bone repair. These preparations
may occur prior to implantation of a solid substrate or simultaneously with implantation.
For example, cartilage and/or bone tissue and/or other tissues proximal to a site
of cartilage and/or bone repair may initially be drilled through to create a channel
of dimensions appropriate for a solid substrate used in the methods of this disclosure.
Then the solid substrate is implanted within the site so that a region of the solid
substrate penetrates the drilled cartilage and/or bone tissues. Alternatively, the
solid substrate may be attached to a tool of this invention capable of penetrating
through cartilage and/or bone or other tissues, or a combination thereof. In this
case, as the tool penetrates through the cartilage and/or bone tissue, the attached
solid substrate is simultaneously implanted.
[0235] In some disclsoures, following implantation of the solid substrate within a repair
site, or several solid substrates within the repair site, the solid substrate is processed
to optimize incorporation and optimal cartilage and/or bone repair. In some disclosures,
such processing may comprise cutting, sanding or otherwise smoothing the surface of
the solid substrate or solid substrates, for optimal repair.
[0236] In one disclosure, methods of this disclosure comprise implanting a solid substrate
in a human subject.
[0237] In one embodiment of this invention, the phrases "long axis of the solid substrate"
and longitudinal axis of the solid substrate" are used interchangeably and refer to
a line extending parallel to the solid substrate lengthwise. The term "lengthwise"
refers the direction of the length of a solid substrate. It may be that an original
geometric shape has been cut to produce a horizontal section of the original solid
substrate. In such instances lengthwise should be viewed as being the original direction
of length along a solid substrate.
[0238] It will be apparent to one skilled in the art that the physical and/or chemical properties
of a solid substrate of this invention and components thereof may influence methods
of use of this disclosure and kits thereof, for inducing or enhancing cartilage and/or
bone repair.
[0239] In one disclosure, methods of this disclosure for inducing or enhancing cartilage
and/or bone repair utilize the 3-D geometry of a solid substrate of this invention
to provide for specifically positioning and confining the solid substrate within a
site of cartilage and/or bone repair.
[0240] One skilled in the art will recognize that the shape of a site of cartilage and/or
bone repair and the shape of a 3-D solid substrate of this invention provide many
different combinations for stably positioning a solid substrate within a site of cartilage
and/or bone repair. In one embodiment, a solid substrate of this invention is shaped
prior to use in methods of this disclosure for cartilage and/or bone repair. In one
embodiment, a solid substrate of this invention is shaped concurrent to use in methods
of this disclosure for cartilage and/or bone repair. By shaping a solid substrate
concurrent with use of the solid substrate in methods of this disclosure, the dimensions
of the solid substrate may be precisely selected for specific positioning of the solid
substrate within a site of repair.
[0241] In one disclosure, methods of this disclosure comprise implanting a solid substrate
in a non-human mammalian and non-mammalian subject. In one embodiment, methods of
this invention comprise implanting a solid substrate in a horse, a race horse, a cow,
a steer, a pig, a rabbit, a goat, a sheep, a farm animal, a pet, a dog, a cat, a monkey,
an ape, a bird and an aves
[0242] In one disclosure, methods of this disclosure are utilized for induced or enhanced
repair of a cartilage and/or bone defect or disorder or disease. In one disclosure,
the cartilage defect results from a trauma, a tear, a sports injury, a full thickness
articular cartilage defect, a joint defect, or a repetitive stresses injury (e.g.,
osteochondral fracture, secondary damage due to cruciate ligament injury). In one
disclosure, the cartilage disorder comprises a disease of the cartilage. In one disclosure,
methods of this disclosure induce or enhance cartilage repair in osteoarthritis, rheumatoid
arthritis, aseptic necrosis, osteochondritis dissecans, articular cartilage injuries,
chondromalacia patella, chondrosarcoma, chondrosarcoma- head and neck, costochondritis
, enchondroma, hallux rigidus, hip labral tear, osteochondritis dissecans, torn meniscus,
relapsing polychondritis, canine arthritis, fourth branchial arch defect or cauliflower
ear. In one disclosure, methods of this disclosure induce or enhance cartilage repair
in degenerative cartilagenous disorders comprising disorders characterized, at least
in part, by degeneration or metabolic derangement of connective tissues of the body,
including not only the joints or related structures, including muscles, bursae (synovial
membrane), tendons, and fibrous tissue, but also the growth plate, meniscal system,
and intervertebral discs.
[0243] In one embodiment, the 3-D shape and chemical composition of a solid substrate of
this invention, used in the methods of this disclosure and/or kits of this invention
will be determined by skilled clinicians, based on factors such as exact nature of
the condition being treated, the severity of the condition, the age and general physical
condition of the subject, body weight, and response of the individual subject, etc.
[0244] In one disclosure, the specific positioning of a solid substrate of this invention
during methods of this disclosure will be determined by skilled clinicians, based
on factors such as exact nature of the condition being treated, the severity of the
condition, the age and general physical condition of the subject, body weight, and
response of the individual subject, etc.
[0245] In one disclosure, methods of this disclosure are evaluated by examining the site
of cartilage and/or bone tissue repair, wherein assessment is by histology, histochemistry,
palpation, biopsy, endoscopy, arthroscopy, or imaging techniques comprising X-ray
photographs, computerized X-ray densitometry, computerized fluorescence densitometry,
CT, MRI or another method known in the art, or any combination thereof.
[0246] In one embodiment, this invention provides a kit for repair of cartilage comprising
the solid substrate of this invention, and directions for utilizing the solid substrate
in cartilage repair.
[0247] One skilled in the art will recognize that choice of a kit by a skilled clinician
would be dependent upon factors such as exact nature of the condition being treated,
the severity of the condition, the age and general physical condition of the subject,
body weight, and response of the individual subject.
[0248] Thus, in one embodiment, the solid substrate comprised in a kit of this invention
comprises different sizes, shapes or chemical compositions, or a combination thereof.
In one embodiment, this invention provides a kit for cartilage repair comprising a
solid substrate of this invention, at least a tool of this invention, and directions
for utilizing the solid substrate in cartilage repair.
[0249] In some embodiments, the term "comprise" or grammatical forms thereof, refers to
the inclusion of the indicated components of this invention, as well as inclusion
of other active agents, and pharmaceutically acceptable carriers, excipients, emollients,
stabilizers, etc., as are known in the pharmaceutical industry.
[0250] In one embodiment, the present invention provides combined preparations. In one embodiment,
the term "a combined preparation" defines especially a "kit of parts" in the sense
that the combination partners as defined above can be used independently or in different
combinations i.e., simultaneously, concurrently, separately or sequentially.
EXAMPLES
EXAMPLE 1
Applications of Coralline-Based Scaffolding of this Invention
[0251] Coralline-based scaffolding of this invention may be inserted into cartilage, bone
or a combination thereof, in a subject in need thereof.
[0252] In some disclosures, such placement will include drilling in the area to expose the
site in which implantation is desired, and tight fitting of the solid substrate within
the defect/site.
[0253] For implantation for cartilage repair, regeneration, etc., solid substrates are implanted
in the desired cartilage site, and within proximally located bone, so that, in this
way, the coral solid substrate is grafted through two types of tissue, cartilage and
bone.
[0254] Figures 1-3 schematically depicts illustrations of embodied solid substrates of this
invention, indicating the presence of the first, second and third phases, respectively
(1-10, 2-10 and 3-10; 1-20, 2-20 and 2-20; and 1-30, 2-30 and 3-30). The series of
hollows 2-40 is evident, as is its impregnation with the second biocompatible polymer
2-45. Figure 4 provides photographs of embodied scaffolds of this invention, with
the three phases indicated, in 4-10, 4-20 and 4-30. Figure 5 schematically depicts
the orientation of a cartoon of a solid substrate of this invention within a site
of cartilage/bone repair.
[0255] Solid substrates may be prepared according to any embodiment as described herein,
as will be appreciated by the skilled artisan.
[0256] The solid substrates are envisioned for use in veterinary applications, as well as
in the treatment of human subjects. It is to be understood that animal studies may
be undertaken to determine optimum configurations and implantation parameters and
procedures.
[0257] For example, animal studies may include implantation of a solid substrate as described
herein within an animal subject and solid substrates are examined and observed over
an extended time period, post surgery. The untreated knee of each animal is used as
a control for comparisons following such surgeries. At appropriate intervals, animals
are sacrificed and histology performed. Appropriate time periods for examining the
site of cartilage repair are 2.5, 4, 9, 12, 26, 52 weeks post surgery. At this time,
the articular surfaces are photographed and tissue is removed from the site of repair
and prepared for histological observations. Specifically, a block consisting of the
grafted area and the surrounding tissue is removed using a fine saw. The material
is further processed for routine histology, which includes slow decalcification.
[0258] In one embodiment of this invention, "about" refers to a quality wherein the means
to satisfy a specific need is met, e.g., the size may be largely but not wholly that
which is specified but it meets the specific need of cartilage repair at a site of
cartilage repair. In one embodiment, "about" refers to being closely or approximate
to, but not exactly. A small margin of error is present. This margin of error would
not exceed plus or minus the same integer value. For instance, about 0.1 micrometers
would mean no lower than 0 but no higher than 0.2. In some embodiments, the term "about"
with regard to a reference value encompasses a deviation from the amount by no more
than 5%, no more than 10% or no more than 20% either above or below the indicated
value.
[0259] In the claims articles such as "a,", "an" and "the" mean one or more than one unless
indicated to the contrary or otherwise evident from the context. Claims or descriptions
that include "or" or "and/or" between members of a group are considered satisfied
if one, more than one, or all of the group members are present in, employed in, or
otherwise relevant to a given product or process unless indicated to the contrary
or otherwise evident from the context. The invention includes embodiments in which
exactly one member of the group is present in, employed in, or otherwise relevant
to a given product or process. The invention also includes embodiments in which more
than one, or all of the group members are present in, employed in, or otherwise relevant
to a given product or process. Furthermore, it is to be understood that the invention
provides, in various embodiments, all variations, combinations, and permutations in
which one or more limitations, elements, clauses, descriptive terms, etc., from one
or more of the listed claims is introduced into another claim dependent on the same
base claim unless otherwise indicated or unless it would be evident to one of ordinary
skill in the art that a contradiction or inconsistency would arise. Where elements
are presented as lists, e.g. in Markush group format or the like, it is to be understood
that each subgroup of the elements is also disclosed, and any element(s) can be removed
from the group. It should be understood that, in general, where the invention, or
aspects of the invention, is/are referred to as comprising particular elements, features,
etc., certain embodiments of the invention or aspects of the invention consist, or
consist essentially of, such elements, features, etc. For purposes of simplicity those
embodiments have not in every case been specifically set forth
in haec verba herein.